8th - University of Manitoba

8th SRCA symposium, May 24-26 2017, Winnipeg

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Welcome

Dear colleagues, friends and attendees,

On behalf of organizing committee for the Society for Research on the Cerebellum

and Ataxia (SRCA) Annual meeting, I am pleased to welcome you to the “8th International

Symposium on the Cerebellum: from Development to Disease”, held in the Rady Faculty

of Health Sciences, University of Manitoba, Winnipeg, May 24-26, 2017.

The SRCA is an international society of scientists and researchers interested in

research on the cerebellum and its associated disorders. In recent years, there has been

tremendous growth in research on cerebellar motor and non-motor functions. The

cerebellum has been shown to play a critical role in diseases ranging from ataxias, autism

spectrum disorders and cognitive operations. The SRCA offers an essential link to

improve, share and intensify this knowledge, by supporting and promoting both basic and

clinical research on the cerebellum. The society’s vision is to promote research and

education, and this symposium provides an excellent platform to fulfill this vision.

I hope you will have three very productive days of interesting and stimulating

discussions. I sincerely wish that this symposium will be a great success not only in

providing an opportunity to share knowledge and expertise in cerebellum research, but

also as the beginning of a long and fruitful cooperation and friendship among fellow

researchers, new investigators and trainees, who will shape our future.

I hope that all of you will enjoy your stay in Winnipeg, one of the “cultural cradles

of Canada,” and Manitoba’s cosmopolitan capital city.

Yours sincerely,

Hassan Marzban

Chair


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Dear colleagues, friends and attendees,

As the president of the Society for Research on the Cerebellum and Ataxia (SRCA)

and on behalf of the Symposium committee members, I am very pleased to welcome you

to the «8th International Symposium on the Cerebellum: from Development to

Disease”, held in Winnipeg, Manitoba, May 24-26, 2017.

This is the 8th meeting of the of the Society for Research on Cerebellum (SRC)

since the inaugural meeting in 2008. It is also the first held under the new name of our

society the Society for Research on Cerebellum and Ataxias (SRCA). In recent years,

there has been tremendous growth in research on cerebellar motor and non-motor

functions, and the cerebellum has been shown to play a critical role in diseases ranging

from ataxias, autism spectrum disorder and cognitive operations. The new title SRCA

reflects the crucial need of interactions between basic scientists and clinicians to increase

our knowledge of cerebellar functions and interactions with other brain regions. This

meeting was organized by Drs. Hassan Marzban and Mario Manto, who along with the

scientific committee, have built a broad and exciting program. Meanwhile, the local

organizing committee has worked to provide the very best conditions for researchers and

trainees to interact, discuss their results and share their hypothesis.

The symposium will be held in the beautiful Basic Medical Sciences Building, on

the Bannatyne campus of the University in Winnipeg. This city offers many attractive

features, from Human Right Museum to Assiniboine Zoo and the famous Winnipeg Ballet,

allowing you to enjoy both excellent science and a pleasant stay.

Once again I welcome you and wish you a wonderful meeting and a nice stay in

Winnipeg

Jean Mariani, President of the SRCA


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Symposium Sponsors

Health Sciences Centre Foundation

Tourism Winnipeg

City of Winnipeg

Department of Human Anatomy and Cell Science

Research Manitoba

University of Manitoba

Max Rady College of Medicine

Rady Faculty of Health Sciences

The Children’s Hospital Foundation University of Manitoba (CHRIM)

Faculty of Graduate Studies

Peter A. Cattini, Henry G. Friesen Chair in Endocrine & Metabolic Disorders


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Department of Biochemistry & Medical Genetics

Department of Pharmacology and Therapeutics

St. Boniface Research Centre

Springer

The Cerebellum


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SFN Cerebellum Social 2017

Tuesday, November 14th, 6:45-8:45pm, Washington D.C.

Chair: Roy V. Sillitoe, PhD, Contact: [email protected]


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PROGRAM:

May 24, 2017

14:00 - 16:00 Registration and Check-in- Brodie Attrium

16:00 - 18:20 Opening Ceremony- Theatre A

16:00 - 17:00 Opening Buffet and Reception- Second Floor Concourse

17:00 - 17:10 Opening: Dr. Hassan Marzban; Chair

17:10 - 17:20 Welcoming Remarks: Dr. Brian Postl; Dean of Medicine

17:20 - 18:20 Session 1: Opening and Plenary Speaker- Theatre A

Session Chair: Dr. Dan Goldowitz

17:20 - 18:20 Dr. Richard Hawkes (CA) -- The Ferdinando Rossi Memorial Lecture: Zones and Stripes - Pattern Formation in the Cerebellar Cortex

18:20 - 21:00 Gala Evening- Brodie Attrium

May 25, 2017

08:45 - 10:25 Session 2: Neuro- and –Morphogenesis- Theatre A

Session Chair: Dr. Michisuke Yuzaki and Dr. Giacomo Consalez

08:45 - 09:10 Dr. Mikio Hoshino (JP) -- Multiple functions of Myeloid Ectopic viral Integration Site 1 homolog in cerebellar granule cell development.

09:10 - 09:35 Dr. Richard Wingate (UK) -- The development of Cerebellar Output

09:35 - 10:00 Dr. Alexandra Joyner (US) -- Cellular interactions underlying proportional scaling of cell types during cerebellar development and repair

10:00 - 10:25 Dr. Joanna Yeung (CA) -- Rhombic Lip Development, Molecular Determinant of Patterning and Cell Specification

10:25 - 10:40 Tea/Coffee- Second Floor Concourse

10:40 - 12:20 Session 3: Normal and Abnormal Differentiation- Theatre A

Session Chair: Dr. Kathleen Millen and Dr. Nori, Koibuchi

10:40 - 11:05 Dr. Azad Bonni (US) -- Epigenetic Regulation of Cerebellar Circuit Assembly and Function

11:05 - 11:30 Dr. James Li (US) -- Bergmann Glia Development, Genesis and Differentiation

11:30 - 11:55 Dr. David Solecki (US) -- Granule Cell Migration -Polarity, Link to Medulloblastoma

11:55 - 12:20 Dr. Martine Roussel (US) -- Epigenetics Drivers in Pediatric Medulloblastoma

12:20 - 13:30 Lunch- Second Floor Concourse


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13:30 - 15:25 Session 4: Circuitry & Functional Development- Theatre A

Session Chair: Dr. Tim Ebner, Dr. Ray Turner and Dr. Marco Molinari

13:30 - 13:55 Dr. Izumi Sugihara (JP) -- The ansiform lobule (crus I in the rodent cerebellum) is unique in its conformation, axonal connection, striped pattern, evolution and development in the mammalian cerebellum

13:55 - 14:20 Dr. Alanna Watt (CA) -- Transient Developmental Purkinje Cell Axonal Torpedoes in Healthy and Ataxic Mouse Cerebellum

14:20 - 14:45 Dr. Karl Schilling (DE) -- Developmental Migration Of Cerebellar Basket and Stellate Cells: Do Synapses Point the Way?

14:45 - 15:10 Dr. Keiko Muguruma (JP) -- Disease modeling with patient-derived iPS cells


15:25 - 17:05 Session 5: Aberrations of Cerebellar Development and Function: Genetics and Imaging- Theatre A

Session Chair: Dr. Jeremy D. Schmahmann and Dr. Esther Becker

15:25 - 15:50 Dr. Michael Salman (CA) -- Epidemiology of Cerebellar Diseases and Therapeutic Approaches

15:50 - 16:15 Dr. Bill Dobyns (US) -- Canary in the coal mine: the cerebellum as a sentinel for developmental brain disorders

16:15 - 16:40 Dr. Catherine Limperopoulos (US) -- Harnessing the power of advanced MRI to understand the role of early-life cerebellar injury on impaired cerebral-cerebellar function

16:40 - 17:05 Dr. Christopher Gomez (US) -- Therapeutic Interventions (SCA)

16:00 – 21:00 Students Networking-TBA

May 26, 2017

08:45 - 10:15 Session 6: Making Connections/Synaptogenesis- Theatre A

Session Chair: Dr. Masanobu Kano and Dr. Rachel Sherrard

08:45 - 09:10 Dr. Keiji Ibata (JP) -- Time-lapse Imaging of Cbln1 Release from Granule Cell Axons and its Accumulation on Purkinje Cell Dendrites

09:10 - 09:35 Dr. Naofumi Uesaka (JP) -- Roles of retrograde signaling in climbing fiber to Purkinje cell synapse elimination during postnatal cerebellar development

09:35 - 10:00 Dr. Fabrice Ango (FR) -- Synaptogenesis: Guidance Molecules in GABA to Pc synapses

10:00 - 10:15 Dr. Laurence Cathala (FR) -- Cellular mechanism of interneuron synaptic integration in developing cerebellum

10:15 - 12:15 Session 7: Posters- Brodie Attrium plus Coffee

Session Chair: Dr. Egidio D'Angelo and Dr. Ying Shen

12:15 - 13:30 plus lunch with experts- Second Floor Concourse


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13:30 - 15:15 Session 8: Aberrations of Cerebellar Development and Function: Motor and Treatment Models- Theatre A

Session Chair: Dr. Roy Sillitoe and Dr. Mario Manto

13:30 - 13:55 Dr. Hirokasu Hirai (JP) -- Regulation of cerebellar function by protein kinase C

13:55 - 14:20 Ms. Lauren Miterko (US) -- Persistent motor dysfunction despite homeostatic rescue of cerebellar morphogenesis

14:20 - 14:45 Dr. Roger Reeves (US) -- Attenuated Shh response in the developing cerebellum of trisomic mice


15:15 - 16:55 Session 9: Autism Spectrum disorder (ASD)- Theatre A

Session Chair: Dr. Bing-wen Soong and Dr. Nicolas Dupré

15:15 - 15:40 Dr. Peter Tsai (US) -- Cerebellar Contribution to Autistic Behaviors

15:40 - 16:05 Dr. Christian Hansel (US) -- Purkinje cell function in mouse models of ASD

16:05 - 16:30 Dr. John Welsh (US) -- ASD and Eyeblink Conditioning

16:30 - 16:55 Dr. Aleksandra Badura (NL) -- Lobule-specific contribution to executive functions in mice

16:55 - 18:00 Closing and AWARD Announcement (Dr. Jean Mariani)


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Speakers

Dr. Richard Hawkes (CA)

The Ferdinando Rossi Memorial Lecture: Zones and Stripes - Pattern Formation in the Cerebellar Cortex. Richard Hawkes, Department of Cell Biology and Anatomy and Hotchkiss Brain Institute, University of Calgary, Calgary, Alberta, Canada.

The cerebellum has a complex architecture – highly reproducible and conserved through evolution. The lecture will first review the molecular patterning of the adult cerebellar cortex and then survey the processes that lead to pattern formation during embryonic development.

Cerebellar architecture is organized around the Purkinje cell. Purkinje cells in the

mouse cerebellum come in many different subtypes, identifiable by expression markers,

sensitivity to mutation etc. These are organized first into four or five “transverse zones”,

each of which is further subdivided into hundreds of reproducible “stripes”. This

arrangement serves as the scaffolding to organize afferent topography and restrict the

distribution of excitatory and inhibitory interneurons. The lecture will first review the

molecular patterning of the adult cerebellar cortex and its conservation through evolution.

Secondly, the lecture will briefly survey some of the mechanisms that lead to

pattern formation during cerebellar development. Pattern formation in the cerebellar

cortex is a multistage process that begins early in development with the generation of the

various Purkinje cell subtypes, and matures through the dispersal of Purkinje cell clusters

into stripes. Two developmental processes will be discussed in particular: the

mechanisms that lead to Purkinje cell subtype specification (i.e., how do we make

different kinds of Purkinje cells?), and the role played by Purkinje cell migration in pattern

formation (i.e., how do the Purkinje cells end up in a reproducible array of stripes?).


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Dr. Mikio Hoshino (JP)

Multiple functions of Myeloid Ectopic viral Integration Site 1 homolog in cerebellar

granule cell development.

Mikio Hoshino, Department of Biochemistry and Cellular Biology, National Institute of

Neuroscience, National Center of Neurology and Psychiatry NCNP, Kodaira, Tokyo,

Japan

Myeloid Ectopic viral Integration Site 1 homolog (Meis1) is a transcription factor of

the TALE (Three Amino acid Loop Extension) protein family. Meis1 has been reported to

maintain the undifferentiated state of progenitor cells, including retinal progenitor cells,

olfactory epithelial cells, and hematopoietic stem cells etc. Although Meis1 expression in

the cerebellum, especially in the EGL (Morales and Hatten, 2006), the function of Meis1

in the granule cell (GC) development has not been clarified.

We reveal that Meis1 is required for proper cerebellar structure formation and for

Pax6 transcription in granule cell precursors (GCPs) and GCs. Meis1-Pax6 pathway

upregulates BMP signaling to induce Atoh1 degradation. However, in the outer EGL,

Meis1 binds to Atoh1 to prohibit its degradation; Meis1-Atoh1 complex upregulates Atoh1

transcription in an autoregulatory fashion. Opposing effects of Meis1 on Atoh1 expression

seem to be attributed to the Atoh1 phosphorylation status. This work reveals multiple

functions of Meis1 to coordinate GC differentiation and gives insights into understanding

neuronal development.


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Dr. Richard Wingate (UK)

The development of cerebellar output

Richard Wingate, MRC Centre - Developmental Neurobiology, King's College London,

United Kingdom

A prominent feature of many cerebellar circuits are the nuclei which receive mossy

and climbing fibre inputs as well as being the target of the majority of Purkinje cell

inhibition. Nuclei send long-range connections to other brain regions that allow the

cerebellum to participate in a variety of central nervous system functions. My group has

examined the origins of nuclear projection neurons in both avian and murine models to

try to understand the factors that regulate their specification. Glutamatergic projection

neurons are derivatives of the Atoh1 positive precursors of the rhombic lip and specified

as part of a sequence of migratory populations. Each population is characterised by a

distinct set of axonal targets. By contrast, inhibitory output neurons, which project only to

the inferior olive, arise from a Sox14 positive pool of precursors that are likely derivatives

of the ventricular zone.


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Dr. Alexandra Joyner (US)

Cellular interactions underlying proportional scaling of cell types during cerebellar

development and repair

Joyner, A.L.1, Wojcinski, A.1, Willet, R.1, Bayin, N.S.1 1Developmental Biology Program, Sloan Kettering Institute, New York, NY, USA

[email protected]

A fascinating question is how during development the number of each cell type in

the cerebellum is generated in the correct proportions (scaled) in order to produce robust

functional circuits. The Purkinje cells have been found to be a critical cell type in the

cerebellar cortex that regulates the expansion of the number of granule cell precursors,

as well as progenitors for interneurons, astrocytes and Bergmann glia produced after

birth. Sonic hedgehog (SHH) secreted by Purkinje cells is the key ligand that regulates

proliferation of the various cortical progenitors after birth. We are exploring the

interactions that occur between the deep cerebellar nuclei, which are the first neurons

born in the cerebellum, and the Purkinje cells that project to the cerebellar nuclei during

embryonic development, by analyzing the phenotypes of mouse engrailed gene (En1/2)

condition mutants. We are characterizing the defects in cerebellar growth resulting from

deletion of En1/2 only in the cerebellar nuclei projection neurons or in the granule cell

precursors, compared to in both cell types generated by the rhombic lip. In another set of

studies, we are testing the degree to which the developing cerebellum can recover from

loss of granule cells soon after birth. The signaling pathways driving normal growth and

recovery after injury have implications for normal development and disease states, as

well as for therapeutic approaches, especially given that the cerebellum is prone to injury

in premature babies, and this is a high risk factor for autism.

mailto:[email protected]


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Dr. Joanna Yeung (CA)

Rhombic Lip Development, Molecular Determinant of Patterning and Cell

Specification

Joanna Yeung, Medical Genetics, University of British Columbia, Vancouver, BC, Canada

The cerebellar rhombic lip (RL) generates all glutamatergic neurons in the

cerebellum. We characterized a novel RL marker, Wntless (Wls), relative to its interaction

with other RL markers (Atoh1, Pax6 and Lmx1a). Using the Wls marker, four distinct

molecular compartments were identified in the developing RL. Our study of Pax6

indicates that Wls is regulated by Pax6 in the RL. Wls is aberrantly expressed in the Pax6

mutant. We also find that the cerebellar nuclear neurons and unipolar brush cells are

missing in the Pax6-null cerebellum, which indicates a novel and crucial role of Pax6 in

the development of all glutamatergic cerebellar neurons. Conversely, the examination of

Wls conditional knockout revealed that Wls regulates Pax6 expression in the RL. Wls

impacts the placement of neurons that leads to an array of ectopic neurons in the Wls-

null cerebellum. Our work demonstrates a novel molecule engaged in cerebellar

development that points to a highly dynamic molecular regulation in the RL during

development.


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Dr. Azad Bonni (US)

Epigenetic Regulation of Cerebellar Circuit Assembly and Function

Azad Bonni, Department of Neuroscience, McDonnell Center for Cellular and Molecular

Neurobiology, Washington University School of Medicine, St. Louis, MO, USA

The chief goal of research in our laboratory is to identify the principles and

mechanisms that govern the assembly and function of neural circuits in the cerebellum

and determine how these mechanisms are deregulated in autism spectrum disorders. We

have discovered fundamental epigenetic, transcriptional, and ubiquitin-signaling networks

that orchestrate distinct aspects of neuronal connectivity in the mammalian cerebellar

cortex. In recent studies, we have identified crucial roles for the nucleosome remodeling

and deacetylase (NuRD) complex in the control of granule neuron parallel fiber

presynaptic differentiation as well as granule neuron dendrite pruning in the mouse

cerebellar cortex in vivo. The NuRD complex triggers long-term silencing of

developmental genes through alterations of histone tail modifications to promote parallel

fiber presynaptic differentiation. By contrast, the NuRD complex dynamically shuts off

activity-dependent gene expression via deposition of the histone variant H2A.z at

promoters of activity-dependent genes to promote granule neuron dendrite pruning and

sparse encoding to sensorimotor stimuli. These findings suggest that the NuRD complex

employs distinct mechanisms to control key aspects of neuronal connectivity in the

cerebellar cortex.


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Dr. James Li (US)

Bergmann Glia Development, Genesis and Differentiation

James Li ,Department of Genetics and Genome Sciences, UConn Health, CT, USA

Folding of the cortex and persistence of radial glia-like cells called Bergmann glia

(BG) are hallmarks of the mammalian cerebellum. Similar to basal radial glia in the

primate neocortex, BG maintain basal processes and molecular features of neural

progenitors. The generation of BG and their role in cerebellar foliation are not well

understood. We have performed mouse genetic experiments, RNA-sequencing, and co-

expression network analyses to study the developmental programs underlying BG

formation. We found that heightened FGF-ERK signaling activity was linked to the timely

transition of radial glia in the cerebellar ventricular zone to BG. Inhibition of FGF-ERK

signaling by deleting Shp2 blocked generation of BG, as well as cerebellar foliation.

Restoring ERK or Etv5 function rescued BG formation in the absence of Shp2. Our results

demonstrate that an FGF-ERK-ETV axis is crucial to BG induction. Furthermore, we

reveal a crucial function of BG in organizing cerebellar foliation.


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Dr. David Solecki (US)

Granule Cell Migration -Polarity, Link to Medulloblastoma

David Solecki, Developmental Neurobiology Department, St. Jude Children's Research

Hospital, TN, USA

Cell polarity is a driving force that coordinates the choreography of neural

development. How polarity signaling organizes the behavior of immature neurons and

how polarity signaling cascades are regulated remain key questions facing the field of

developmental neurobiology. These questions are critical to understand the pathology of

neurodevelopmental diseases, where the production of neurons or their subsequent

migration is defective. Studies combining necessity-sufficiency testing and cutting edge

imaging technology in the developing cerebellum show that a conserved polarity-signaling

module, called the Pard complex, is essential for neuronal progenitor germinal zone exit

by regulating cytoskeletal dynamics and cell-cell interactions needed for neuronal

migration. I will present our progress identifying an upstream regulator of the Pard

complex: an E3 ubiquitin ligase, Seven in Absentia, which mediates proteosomal

degradation of Pard3; to control a shift from tangential to radial migration when cerebellar

granule neurons leave their mitogenic niche, and drebrin; to control microtubule-actin

crosslinking during CGN differentiation.


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Dr. Martine Roussel (US)

Epigenetic Drivers in Pediatric Medulloblastoma

Martine Roussel, Department of Molecular Sciences, St. Jude Children's Research

Hospital, TN, USA

-Medulloblastoma (MB), an embryonal cerebellar tumor, comprises four distinct

subgroups: Sonic Hedgehog (SHH), Wingless (WNT), Group3 (G3) and Group4 (G4).

MBs have a paucity of mutations with genetic alterations in oncogenes and tumor

suppressors (β-CATENIN, PATCHED, SUFU, GLI2, MYC and MYCN) account for

only 20-30% of cases. Next generation sequencing revealed somatic altered genes,

many of which involved in epigenetic regulators and chromatin modification.

Studies by the Washington University/ St. Jude Pediatric Cancer Genome Project

showed that mouse and human G3 MBs express increased levels of EZH2, the

catalytic partner of the polycomb repressor complex PRC2, and of histone 3 lysine

27 trimethylation (H3K27me3). Deletion of EZH2 or SUZ12 in G3 MB via TALEN and

CRISPR-Cas9 gene editing approaches revealed that the PRC2 complex has tumor

suppressive functions in G3 MB, via in part the suppression of GFI1, a transcriptional

repressor overexpressed in a subset of G3 MBs.


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Dr. Izumi Sugihara (JP)

The ansiform lobule (crus I in the rodent cerebellum) is unique in its conformation,

axonal connection, striped pattern, evolution and development in the mammalian

cerebellum

Izumi Sugihara, Dept. Systems Neurophysiol Tokyo Medical and Dental Univ., Tokyo,

Japan

In the human cerebellum, crus I and crus II lobules (or ansiform lobule), which are

implicated in cognitive and visuomotor functions, are significantly expanded compared to

anterior and posterior lobules, which are involved in somatosensorimotor function. In

applying rodent models, it is essential to identify the lobules that are homologous to

human crus I and crus II. Observation of the lobular structure in human, macaque,

marmoset, rat, and mouse indicated that the human crus I/II were homologous to

crus I in rodents (referred to as “ansiform area, AA”). Our lobular definition was

supported by lobule-based mapping of the olivocerebellar climbing fiber and Purkinje cell

(PC) projection patterns in rodents; Crus II and simple lobule was innervated by the

mediocaudal part of each inferior olive subnucleus and project to the dorsal part of

the cerebellar nuclei, while crus I (or the AA) was innervated by the rostrolateral part

of each inferior olive subnucleus and project to the ventral part of the cerebellar nuclei.

A gap in the cortical structure was observed in the paravermal area of the AA in both

rodents and primates. Concerning zebrin stripes, the central lobules (lobules VI-VII, and

AA or crus I in rodents) show a laterally-expanded arrangement solely of positive stripes.

Our analysis showed that this arrangement of zebrin-positive stripes in the AA originated

from their developmental process. Between E14.5 and E17.5, lateral protrusion and shift

were observed in the domains of protocadherin 10-positive PC subsets (which would

become zebrin-positive later) in the central area of the immature cerebellum that would

eventually become lobules VI-VII and AA or crus I. The results indicate that the AA (or

crus I in rodents) is characterized by distinct connectivity from neighboring lobules and a

massive expansion in skillful primates.


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Dr. Alanna Watt (CA)

Transient Developmental Purkinje Cell Axonal Torpedoes in Healthy and Ataxic

Mouse Cerebellum

Alanna Watt, Department of Biology, McGill University, Montreal, Quebec, CA

Information is carried out of the cerebellar cortical microcircuit via action potentials

propagated along Purkinje cell axons. In several human neurodegenerative diseases,

focal axonal swellings on Purkinje cells – known as torpedoes – have been associated

with Purkinje cell loss. Interestingly, torpedoes are also reported to appear transiently

during development in rat cerebellum. The function of Purkinje cell axonal torpedoes in

health as well as in disease is poorly understood. We are investigating the properties of

developmental torpedoes in the postnatal mouse cerebellum of wild-type and transgenic

mice. Our findings to date suggest that the transient emergence of Purkinje cell axonal

torpedoes during the second postnatal week in mice represents a normal morphological

feature in the developing cerebellar microcircuit.


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Dr. Karl Schilling (DE)

Developmental migration of cerebellar basket and stellate cells: do synapses

point the way?

Karl Schilling Anatomisches Institut, Anatomie & Zellbiologie Universität Bonn, Bonn,

Germany. [email protected]

In the developing cerebellum, molecular layer inhibitory interneurons are among

the last nerve cells to reach their adult positions and to differentiate. They originate from

the ventricular epithelium lining the fourth ventricle, and their distribution to and within the

cerebellar cortex requires extensive migration through dynamically changing cellular

environments, from the nascent white matter through the immature granule cell layer, and

finally within the emerging molecular layer. The mode of migration of these cells, the cues

they use to navigate, and the mechanisms to interpret such cues remain largely elusive.

I will summarize some recent data that allows us to describe and to quantify how

molecular layer interneurons navigate through the nascent cerebellar cortex. Further, I

will present data that show that these cells are synaptically innervated much earlier than

hitherto thought, and in fact while still in transit. This developmental vesicular transmitter

release is part of the machinery that ensures proper migration and navigation of these

cells. Beyond the implications for our understanding of cerebellar histogenesis, these

findings suggest a novel mechanism how functional activity of early maturing nerve cells

may tune the cellular composition and functional properties of emergent nerve cell

networks. These findings also define a hitherto unknown role for early synapses as

activity-tunable guideposts for neural migration.



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Dr. Keiko Muguruma (JP)

Disease modeling with patient-derived iPS cells

Keiko Muguruma, RIKEN, Center for Developmental Biology (CDB), Laboratory for Cell

Asymmetry, 2-2-3 Minatojima-minamimachi, Chuo, Kobe, Japan

Recent advances in the techniques that differentiate induced pluripotent stem cells

(iPSCs) into specific types of cells enabled us to establish in vitro disease models from

the patient-derived iPSCs. The advantage of the model utilizing disease-specific iPSC

is that it is able to generate a large number of cells required for high-throughput

screening. The patient-derived iPSCs are expected to recapitulate the disease-specific

pathogenesis and physiology in vitro. We developed 3D culture systems using human

pluripotent stem cells, which would promote the research on the construction of complex

brain regions. Recently we have succeeded in generation of spinocerebellar ataxia

(SCA), SCA6 and SCA42, patient-derived Purkinje cells by combining the iPSC

technology and the self-organizing stem cell 3D culture technology. We have constructed

an in vitro disease model recapitulating both ontogenesis and pathogenesis for SCA.

Here we will talk about approaches for intractable diseases utilizing patient-specific

iPSCs.


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Dr. Michael Salman (CA)

Epidemiology of cerebellar diseases and therapeutic approaches

Michael Salman, Department of Pediatrics and Child Health, University of Manitoba,

Winnipeg, MB, Canada

Cerebellar diseases occur relatively commonly in children and adults around the

globe. Many factors influence their epidemiology including geography, ethnicity,

consanguinity, and the methodology used to ascertain patients. In addition, reliable

epidemiological data relies heavily on accurate disease classification. The continuous

advances in genetics and neuroimaging modalities have resulted in improved

understanding of cerebellar diseases and have led to several revisions in their

classification. Recent global epidemiological studies on ataxia reported an estimated

overall prevalence rate of 26/ 100,000 in children, a prevalence rate of dominant

hereditary cerebellar ataxia of 2.7/ 100,000, and a prevalence rate of recessive hereditary

cerebellar ataxia of 3.3/ 100,000. The management of cerebellar diseases is

multidisciplinary and multimodal. General supportive and symptomatic therapies should

be initiated. Genetic counselling should be offered, where appropriate. Few drugs,

specific motor rehabilitation programs, and non-invasive cerebellar stimulation for the

treatment of ataxia have been developed and seem to show early promise but more

studies are needed to replicate and fine-tune their benefits further. Some disease-specific

treatments are available. For example, acetazolamide or 4-aminopyridine for patients with

episodic ataxia type 2 and vitamin E for patients with vitamin E deficiency.


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Dr. Bill Dobyns (US)

Canary in the coal mine: the cerebellum as a sentinel for developmental brain

disorders

Dobyns WB1,2; 1Center for Integrative Brain Research, Seattle Children’s Research

Institute, Seattle, WA, USA, 2Department of Pediatrics, University of Washington, Seattle,

WA, USA

The cerebellum is often overlooked in assessing fetuses and children with

developmental brain disorders because of multiple patterns of malformation that are

inconsistently defined, lack of experience in recognizing these patterns, variable severity

including non-penetrance, occasional co-occurrence of atrophy, and limited

understanding of the underlying causes. After excluding two well-known groups of

autosomal recessive disorders with recognizable patterns of malformation (Joubert

syndrome and pontocerebellar hypoplasia), cerebellar malformations have been

consistently observed with only two copy number variants (deletion 3q24 or 6p25.3) and

a few genes (CASK, OPHN1 and FOXC1). Recent experience has shown that prenatal

events such as late 2nd-early 3rd trimester posterior fossa and cerebellar bleeds and

(less often) cerebellar ischemia, can cause cerebellar injuries that mimic cerebellar

malformations. Accordingly, genetic studies have shown a lower rate of abnormalities

than other developmental disorders such as agenesis of the corpus callosum, intellectual

disability and autism.

We have performed SNP microarrays in ~250 children and whole exome

sequencing data in ~100 children (and parents) with cerebellar malformations. Our

analysis suggests that cerebellar hypoplasia is a variable feature in many genetic

developmental brain disorders, that prenatal injuries to the cerebellum are common and

can often be recognized based on the pattern of abnormality, and that these two

processes may co-occur.


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Dr. Catherine Limperopoulos (US)

Harnessing the power of advanced MRI to understand the role of early-life

cerebellar injury on impaired cerebral-cerebellar function.

Catherine Limperopoulos, PhD

Director, the Developing Brain Research Program Vice Chief of Research, Division of

Diagnostic Imaging and Radiology

Co-Director of Research, Division of Neonatology Children’s National Health System

Associate Professor of Neurology, Radiology, and Pediatrics George Washington

University School of Medicine and Health Sciences

Cerebellar development follows a highly orchestrated and complex program of

critical developmental processes. Consequently, this vulnerable developmental period

of the cerebellum can be derailed by a host of potential insults. Recently, we have applied

quantitative MRI (qMRI) tools to study the developmental trajectory of the human

cerebellum in utero. Using qMRI we have demonstrated that the cerebellum undergoes

its most rapid growth that is unmatched by any other cerebral structure over the third

trimester. However, this accelerated growth is impeded by premature birth where many

of these complex cerebellar development events take place within the hazards of a hostile

extrauterine environment. We will review the role of both direct and indirect cerebellar

injury on cerebral development and relate these disturbances in cerebellar development

to a prevalent and distinct profile of cognitive, language and social-behavioral dysfunction

including autism spectrum disorders. Finally, we will explore the emerging functional

topography of the immature cerebellum and its relationship to long-term

neurodevelopmental disabilities.


28

Dr. Christopher Gomez (US)

Therapeutic Interventions (SCA)

Department of Neurology, The University of Chicago, Chicago IL, USA

We have discovered that the P/Q-type voltage-gated Ca2+ channel (VGCC) gene,

CACNA1A, is a bicistronic cellular gene, i.e, encodes two structurally unrelated proteins,

with distinct functions, that are separately encoded within the same mRNA. CACNA1A

encodes both the α1A (Cav2.1) subunit and a newly recognized transcription factor,

α1ACT, within an overlapping open reading frame (ORF) within the same mRNA

transcript. This is achieved by the presence of a novel internal ribosomal entry site (IRES)

upstream of a second ORF encoding α1ACT, which, when mutated, mediates the

disease, spinocerebellar ataxia type 6 (SCA6). The IRES controlling α1ACT is an

excellent drugable target and we have used this strategy to suppress an acute form of

SCA6. However, this approach suppresses both the normal and expanded α1ACT.

Therefore, we have used a doxycycline suppressible wild type α1ACT transgene to

demonstrate its critical role in early Purkinje cell maturation, whose elimination in adults

does not have adverse effects on cerebellar cortex function.


29

Dr. Keiji Ibata (JP)

Time-lapse imaging of Cbln1 release from granule cell axons and its accumulation

on Purkinje cell dendrites

Ibata K1, Yuzaki M1; 1Department of Physiology, Keio University School of Medicine

[email protected]

Cbln1 belongs to the C1q/TNF superfamily, whose members are involved in

various intercellular signaling in multiple systems. Cbln1 is released from cerebellar

granule cells and plays an essential role in formation and maintenance of synapses

between parallel fibers and Purkinje cells (Matsuda et al, Science 2010). Cbln1 forms a

tripartite complex across the synapse by binding to its pre- and postsynaptic receptors,

neurexin (Nrx) and the delta2 glutamate receptor (GluD2). Nevertheless, a fundamental

question how and where Cbln1 is released from granule cells has remained unclear. To

perform time-lapse imaging of Cbln1 release, we expressed Cbln1 tagged with pH-

sensitive GFP (super-ecliptic-pHluorin; SEP) in cultured granule cells. Electrical field

stimulation rapidly increased SEP-Cbln1 fluorescence signals from mostly non-synaptic

sites, which were negative for presynaptic markers. Upon encountering with HEK293 cells

expressing GluD2, surface SEP-Cbln1 signals along granule cell axons started to

accumulate at the contact site in the absence of electrical field stimulation. From these

results, we propose that Cbln1, secreted from extrasynaptic sites in an activity-dependent

manner, accumulates on the region where axons make contact with GluD2 on dendritic

spines of Purkinje cells in an activity-independent manner.



30

Dr. Naofumi Uesaka (JP)

Roles of retrograde signaling in climbing fiber to Purkinje cell synapse

elimination during postnatal cerebellar development

Uesaka N1, Abe M2, Yamazaki M2, Konno K3, Mikuni T1, Watanabe M3, Sakimura

K2and Kano M1 1The University of Tokyo, Tokyo, Japan 2Niigata University, Niigata, Japan 3Hokkaido University, Sapporo, Japan

Presenting author’s e-mail address: [email protected]

Purkinje cells (PCs) in the neonatal cerebellum are innervated by multiple climbing

fibers (CFs). During postnatal development, a single CF is selectively strengthened in

each PC and becomes a ‘winner’ CF that is presumed to remain into adulthood, whereas

the other ‘loser’ CFs are eliminated. Our recent studies have uncovered molecular

mechanisms by which postsynaptic PCs regulate CF synapse elimination. We have

demonstrated that Semaphorin7A and Semaphorin3A mediate retrograde signals from

postsynaptic PCs to presynaptic CFs and regulate elimination and maintenance

respectively of CF synapses. We further screened candidate molecules that may mediate

retrograde signaling for strengthening or weakening of CF synapses. We found that PC-

specific deletion of progranulin, a growth factor implicated in the pathogenesis of

frontotemporal dementia, and knockdown of Sort1, a receptor of progranulin, in CFs

accelerated elimination of redundant CFs and reduced the amplitude of synaptic inputs

from winner CFs. These results suggest that progranulin derived from PCs retrogradely

acts on Sort1 in CFs, strengthens/maintains CF synapses.


31

Dr. Fabrice Ango (FR)

Synaptogenesis: Guidance Molecules in GABA to Pc synapses

Fabrice Ango, IGF/Neurobiology, Université de Montpellier, France

One of the remarkable features of neuronal circuits is the specificity and precision

of the synaptic connections during development. These highly specific patterns of

connections between different populations of neurons are crucial for brain function, and

require an intricate coordination of various developmental events. However, little is known

about how growing axons select correct post-synaptic partner at the cellular and

subcellular level within multiple heterogeneous targets they encounter. This is well

exemplified by GABAergic interneurons, which innervated specific cell types at precise

subcellular location (i.e. dendrites, cell soma or axon initial segment (AIS)). Our recent

studies suggest that the axon guidance receptor Neuropilin-1 expressed by GABAergic

interneurons orchestrated both guidance and subcellular synapse targeting through

timely interactions with local cues. Coordination of both guidance and recognition by the

same molecular cue might alleviate some of the coding power for synapse specificity.


32

Dr. Laurence Cathala (FR)

Cellular mechanism of interneuron synaptic integration in developing cerebellum

Laurence Cathala; Adaptation Biologique et Vieillissement, CNRS UMR 8256, Case 14,

Université Pierre et Marie Curie – P6, Sorbonne Universités, 9 Quai St Bernard, 75005

Paris, France. [email protected]

Interneurons are critical for neuronal circuit function, but how their dendritic

morphologies and membrane properties influence information flow within neuronal

circuits is not well understood, let alone how these properties change with development

and allow networks to acquire and refine their functional properties. We have addressed

this question by studying synaptic integration of excitatory inputs onto stellate cells,

molecular layer interneurons, in the immature cerebellum. With a multidisciplinary

approach combining electrophysiological recording in brain slices, morphological analysis

and neuronal simulation, we found that information processing from immature stellate

cells differs from what we previously described in the adult stellate cell (Abrahamsson et

al., 2012). This developmental change in the integration of excitatory synaptic inputs

results predominantly from a difference in synapse location and quantal size. These

alterations are likely to lead to a shift in the subthreshold input-output transformations

from more linear relationship in young SCs to a more sublinear relationship in older SCs.



33

Dr. Hirokazu Hirai (JP)

Regulation of cerebellar function by protein kinase C

Hirokazu Hirai, Department of Neurophysiology & Neural Repair, Gunma University

Graduate School of Medicine

Prof. Yasutomi Nishizuka discovered protein kinase C (PKC), a family of serine-

and threonine-specific protein kinases, which regulates myriad of physiological functions.

I learned biochemistry including PKC from Prof. Nishizuka in Kobe University just around

the time he and his lab members were actively studying the mechanism of PKC activation

following membrane lipid hydrolysis. The PKC that Prof. Nishizuka originally discovered

is categorized in classical (conventional) PKCs. Classical PKCs (PKCα, PKCβI/βII and

PKCγ) are activated by calcium and second messenger diacylglycerol, in which PKCγ is

expressed exclusively in neurons of the brain and spinal cord. Cerebellar Purkinje cell

(PC) expresses PKCα and PKCγ. PKCα is indispensable for the expression of cerebellar

long-term depression (LTD) at parallel fiber to PC synapses because PKCα can bind to

PICK1 via the PDZ domain-binding motif, accesses to and phosphorylates the C-terminal

domain of GluA2, eventually leading to clathrin-mediated endocytosis of postsynaptic

AMPA receptors. Systemic PKCγ-knockout mice have been shown to exhibit deficient

pruning of climbing fibers (CFs) from developing PCs and mild motor coordination deficit,

suggesting a critical role in elimination of CF synapses from PCs during development.

There are currently, at least, 2 open questions. First one is why PKCα cannot substitute

for PKCγ to eliminate surplus CFs from developing PKCγ-null PCs. Second one is what

role PKCγ plays in after maturation of the cerebellum, which has remained totally

unknown. We challenged to resolve these 2 questions.


34

Ms. Lauren Miterko (US)

Persistent motor dysfunction despite homeostatic rescue of cerebellar

morphogenesis

Miterko LN and Sillitoe RV; Pathology & Immunology, Neuroscience, Developmental

Biology, Baylor College of Medicine, Duncan Neurological Research Institute, Houston

Texas 77030, USA

Purkinje cells play a central role in establishing the cerebellar circuit. It is well

known that disrupting Purkinje cell development impairs cerebellar morphogenesis and

motor function. Surprisingly, in the Car8wdl mouse model of hereditary cerebellar ataxia,

severe motor deficits arise despite the cerebellum developing to its correct size and

morphology. We revealed that loss of the Purkinje cell protein called CAR8 (Carbonic

anhydrase 8), a regulator of IP3R1 Ca2+ signaling, delays cerebellar morphogenesis by

transiently restricting growth. The mechanism involved a reduction of granule cell

proliferation as observed in postnatal day (P) 5 mutants, although by P15 proliferation

was maintained at a higher level compared to controls. The prolonged period of granule

cell proliferation was accompanied by a restructured assembly of Purkinje cell and

Bergmann glia architecture, which both coordinate granule cell migration. We next used

in vivo electrophysiology, EMG, and behavior to show that the onset of motor dysfunction

occurs by P20. Together, our findings indicate that CAR8 mediates Purkinje cell-granule

cell communication during cerebellar growth, and its loss triggers an inter-cellular

compensatory response to rescue structure, but not motor function. These data raised

the possibility of using the Car8wdl model to test whether therapeutic targeting of an adult

circuit can be used to correct a developmentally derived behavioral deficit. Towards this,

we devised a cerebellar nuclei deep brain stimulation (DBS) approach to correct

movement in mice. After four days of DBS treatment, Car8wdl mutant mice showed

sustained increases in motor performance and learning. Given these data, we propose

that cerebellar DBS could be a promising therapy for non-degenerative cerebellar ataxias

and in addition Car8wdl mice may provide a critical new opportunity to finally solve the

cellular mechanism(s) for how DBS works in vivo.


35

Dr. Roger Reeves (US)

Attenuated Shh response in the developing cerebellum of trisomic mice

Roger H. Reeves, Johns Hopkins Univ. Schl. Med., Baltimore, MD USA

Trisomy 21 results in Down syndrome which presents as a constellation of features

including a significantly reduced volume and cellularity of the cerebellum. This reduction

also occurs in mouse models of Down syndrome such as Ts65Dn, which is trisomic for

orthologs of more than half of the genes conserved with human chromosome 21 (Hsa21).

We traced the basis of the defect to the proliferation of granule cell neuron precursors

(gcp) in the days after birth. This reduction could be attributed substantially to an

attenuated gcp response to the mitogenic effects of Shh growth factor. A single injection

on the day of birth of the Shh pathway agonist, SAG, normalized cerebellar structure

through life and produced a surprising improvement in hippocampal function in behavior

tests and a normalization of LTP, an electrophysiological correlate of hippocampal

learning. We are pursuing three basic lines of research, one defining the brain regions

where acute Shh pathway stimulation around birth can improve synaptic function in

hippocampus throughout life, a second characterizing anatomical changes in tracts from

deep cerebellar nuclei throughout the brain, and a third seeking to identify the trisomic

gene or genes that are responsible for the attenuated response to Shh.


36

Dr. Peter Tsai (US)

Cerebellar Contribution to Autistic Behaviors

Peter Tsai, Department of Neurology and Neurotherapeutics, Neuroscience, Pediatrics,

Psychiatry, Center for Autism and Developmental Disabilities, University of Texas

Southwestern, TX, USA

The cerebellum has been implicated in the pathogenesis of multiple

neuropsychiatric disorders. In particular, cerebellar abnormalities and pathology has been

identified in many studies of autism while cerebellar disorders predispose to significantly

increased rates of autism. We have recently demonstrated that cerebellar dysfunction is

sufficient to generate autistic behaviors. We will discuss our recent studies investigating

sensitive periods of treatment and mechanisms by which the cerebellum regulates these

behaviors.


37

Dr. Christian Hansel (US)

Purkinje cell function in mouse models of ASD

Christian Hansel (University of Chicago), USA

Autism Spectrum Disorder (ASD) is characterized by deficits in social interaction

and by repetitive behaviors, and is often accompanied by motor impairment. In a mouse

model for the human 15q11-13 duplication (Dup15q syndrome), which often presents

itself with autism, delayed motor milestones and seizures, we have found abnormal

synaptic function and plasticity at cerebellar synapses as well as impaired eyeblink

conditioning (EBC), a form of cerebellum-dependent motor learning that is also affected

in ASD patients. We currently perform studies to determine the cause of motor deficits in

Dup15q syndrome / ASD, and to characterize synaptic and behavioral phenotypes in

CYFIP1 overexpressing mice to assess whether increased dosage of this candidate gene

causes ASD-typical motor and non-motor phenotypes / symptoms in Dup15q syndrome.


38

Dr. John Welsh (US)

ASD and Eyeblink Conditioning

Dept of Pediatrics, Seattle Children's Hospital, University of Washington, Seattle, WA,

USA

The inferior olive (IO) has been implicated in motricity and in associative learning

as measured by Pavlovian conditioning of the eyeblink response. Spike synchrony among

IO neurons, enabled by phase-locked, subthreshold oscillations (STOs) in membrane

potential, is relevant for the fine temporal control of movement and the activation of

muscles as collectives – or as “muscle synergies.” Multiple lines of evidence indicate that

STOs are an emergent property enabled by weak, but prevalent, electrical coupling. We

demonstrated NMDA receptor-mediated strengthening of electrical coupling between IO

neurons that synchronizes STOs and increases their amplitude. I will discuss the

influence of the upward modulation of STO amplitude and synchrony by NMDA-receptor

mediated plasticity of electrical synapses for motricity and associative learning, and its

possible relevance to childhood autism in which there is a range of motor and associative

learning impairments.


39

Dr. Aleksandra Badura (NL)

Lobule-specific contribution to executive functions in mice.

Aleksandra Badura, Netherlands Institute for Neuroscience, Amsterdam Netherlands

Cerebellar lobules VI/VII and crus I/II form reciprocal loops with neocortical regions

associated with executive functions. To test the functional significance of those

connections we used inhibitory DREADDs to disrupt neural activity of these cerebellar

regions during postnatal development or adulthood, and measured the consequences in

two major domains: (1) social choice and behavioral inhibition, as measured using a

three-chamber mouse/object test and an elevated-plus maze; and (2) cognitive flexibility,

as measured by the ability to change the preferred arm in a Y-maze swim task and time

spent on grooming. In lobule VI, cerebellar disruption in adult miceled to increased

perseveration. Mice could learn to find a hidden platform in a swimming Y-maze, but

showed impaired switching when the platform was moved to the other arm of the maze.

The mice also showed reduced performance and bias in a virtual reality-based working

memory task. However, preference for mouse over object in the three-chamber test was

unchanged. These results suggest that lobule VI is an active part of the brain wide circuitry

for cognitive flexibility. Developmental effects were seen from perturbations of crus I, crus

II, and lobule VII. Unilateral, developmental disruption of cerebellar activity in crus I led to

reduced social interaction in a three-chamber test and impaired reversal learning in a Y-

maze, consistent with a role for crus I in social and cognitive maturation. Unilateral crus

II perturbation during development led to reductions in movement in the elevated-plus

maze and three-chamber task, as well as reduced social preference in the three-chamber

test, consistent with a broad role for crus II in the maturation of behavioral inhibition. None

of these effects were seen in the control group. In lobule VII, developmental perturbation

affected exploratory activity in the elevated-plus maze and in the three-chamber test.

Opposite effects were observed in acute, adult cerebellar disruption, suggesting that

lobule VII contributes to maturation and acute function of exploratory behavior. Our results

are consistent with the hypothesis that the cerebellum actively contributes to executive

functions. Furthermore, these experiments provide first direct evidence for the idea that

the cerebellum acts during sensitive periods to shape the developing brain (Wang et al.,

2014).


40

Poster Number/Presenter/Title

Theme 1- Cerebellar Neuro- and Morphogenesis.

P1 / Mrs. Maryam Rahimi-Balaei/ The Role of a Novel Subset of Mesencephalic Neural Crest

Derived Cells in Cerebellar Nuclei Development in Mice

P2 / Miss. Tsz Ching Ma/ Canonical BMP signaling is required to maintain neural stem cells at

cerebellar ventricular zone.

P3 / Prof. Annalisa Buffo/ Multiple origins and spatiotemporal emergence of cerebellar astrocyte

heterogeneity.

P4 / Dr. Filippo Casoni/ Zfp423, a Joubert syndrome gene, is a domain-specific regulator of cell

cycle progression, DNA damage response and Purkinje cell development in the cerebellar

primordium.

P5 / Prof. Im Joo Rhyu/ Stereological analysis of Purkinje cell synapse in the molecular layer of

the rat cerebellum according to its phylogenic lobules.

P6 / Dr. Daniel Turnbull/ In Vivo 4D MRI of the Developing Mouse Cerebellum.

P7 / Dr. Parthiv Haldipur/ Disrupted rhombic lip development caused by aberrant mesenchymal

signaling likely represents a unifying developmental mechanism for human Dandy-Walker

malformation.

P8 / Dr. Thomas J. Ha/ Identification of key regulators for cerebellar development using

FANTOM5 time-course CAGE data.


41

P1

The Role of a Novel Subset of Mesencephalic Neural Crest Derived Cells in Cerebellar

Nuclei Development in Mice

Rahimi Balaei M 1, Ashtari N 1, Jiao X 1, Hassan Marzban 1

1Department of Human Anatomy and Cell Science, The Children’s Hospital Research

Institute of Manitoba (CHRIM), Max Rady College of Medicine, Rady Faculty of Health

science, University of Manitoba, Winnipeg, MB, R3E 0J9, Canada,

[email protected]

Introduction: During cerebellar development, cerebellar nuclei (CN) neurons and Purkinje

cells are the earliest born among the different neuronal subtypes. Purkinje cells are the

sole output of the cerebellar cortex that project to the CN. The CN represents the main

output of the cerebellum, which is generated from the rhombic lip. In this study, we

investigated new origin for subet of the CN neurons during early cerebellar development.

Methods: We used whole mount/section immunohistochemistry, cerebellar culture,

Western blot, and embryonic cultures to examine the origin of a new subset of CN

neurons from the mesencephalon during early cerebellar development.

Results: Our results show that a subset of CN neurons, which are immunopositive for α-

synuclein (SNCA) and orthodenticle homeobox 2 (Otx2), originate from the

mesencephalon and cross the isthmus toward the rostral end of the nuclear transitory

zone. Interestingly, double immunostaining of the SNCA with Otx2 or p75 neurotrophin

receptor (p75ntr) suggests that these cells are probably derived from neural crest cells.

We also showed that this population of neurons with nerve fibers terminates at the subpial

surface of putative lobules VI/VII. The SNCA+/Otx2+/p75+ cells, which divide the

cerebellar primordium into rosterodorsal and caudoventral compartments, show

increased cleaved caspase-3 (CC3+) activation.

Conclusion: These results suggest that early CN neurons originate from the

mesencephalic neural crest population; contrary to popular opinion that Otx2 has been

shown to be involved in prosencephalon and mesencephalon establishment, but not the

rhombencephlon. The p75 immunopositive cells which show activation of caspase-3

during embryonic stage suggest their role in proliferation, differentiation, survival and

axonal guidance. The presence of migratory mesencephalic derived neural crest cells in

the nuclear transitory zone suggests that these neurons/fibers have a regulatory role as

a signaling center that may play as an intrinsic organizer during early cerebellar

development.


42

P2

Canonical BMP signaling is required to maintain neural stem cells at cerebellar

ventricular zone

Ma, T.C.1, Vong, K.I.1, Kwan, K.M.1, 2, 3

1 School of Life Sciences, 2 Centre for Cell and Developmental Biology, 3 State Key

Laboratory of Agrobiotechnology (CUHK), The Chinese University of Hong Kong, Hong

Kong, China.

The anterior rhombic lip (ARL) and ventricular zone (VZ) are functionally distinct neural

stem cell pools in embryonic cerebellum. While ARL generates glutamatergic neurons,

cerebellar VZ is responsible for the production of GABAergic neurons and glial cells.

Canonical BMP signaling is essential to cell specification at ARL, while its role at

cerebellar VZ remains largely unknown. In view of the expression of phosphorylated

Smad1/5 in VZ at embryonic day (E) 11.5 of mouse, we hypothesize canonical BMP

signaling regulates neural stem cell maintenance and/or neurogenesis at cerebellar VZ.

We found that conditional knockout of Smad1/5 via Engrailed 1 (En1) promoter-driven

Cre resulted in drastic reduction in cell proliferation at cerebellar VZ. To assess the

depletion rate of neural stem cells, we examined the expression pattern of Sox2, a neural

stem cell marker, by immunohistochemistry. Our results revealed a quicker depletion of

neural stem cells at the VZ in mutant cerebella. Loss of Smad1/5 promoted specification

of neural stem cells at cerebellar VZ and this led to increased neurogenesis. On the other

hand, radial glial cells at cerebellar VZ give rise to Bergmann glia and astrocytes from

around E14. Therefore, we analyzed the expression pattern of radial glial cells/Bergmann

glia marker BLBP and astrocyte marker GFAP. Our results suggested generation of

Bergmann glia was also impaired in Smad1/5 mutants. Taken together, canonical BMP

signaling plays a crucial role in neural stem cell maintenance at cerebellar VZ. Smad1/5

is required to prevent premature neurogenesis and enables proper development of

Bergmann glia.


43

P3

Multiple origins and spatiotemporal emergence of cerebellar astrocyte heterogeneity

Cerrato V. 1, Parmigiani E.1, Figueres Oñate M.2, de’Sperati C.4, Lopez-Mascaraque

L.2, Buffo A.1

University of Turin

Despite astrocytes are viewed as a homogeneous population, a growing body of evidence

indicates a high degree of morphological, molecular and functional astroglial

heterogeneity. Yet, the developmental processes that lead to this heterogeneity are still

unclear. The cerebellum, with its variety of morphologically distinct astroglial phenotypes

allocated in different layers, is an excellent model to address this issue. To this aim, we

performed in vivo clonal analysis of embryonic ventricular progenitors using Star Track

plasmids. Clone dispersion revealed that astrocyte generation follows the spatiotemporal

pattern of birth of Purkinje neurons, with early and late-generated clones being located in

the most lateral or medial parts of the cerebellum, respectively. Further analyses

disclosed the existence of four major ventricular progenitor types producing either

granular layer (GL) or white matter (WM) astrocytes, or mixed progenies including

Bergmann glia (BG) and GL astrocytes or all types of cerebellar astrocytes. Notably,

postnatally radial progenitors in the Purkinje cell layer that divide in situ to generate both

BG and GL. Moreover, the frequency of mixed progenies declines with time together with

clone size and spatial dispersion, indicating a time-regulated decrease in fate potential.

Interestingly, triple clones showed a constant architecture, with astrocytes in the cortical

layers outnumbering those in the WM. This suggests that layer-specific dynamics regulate

the amplification of sister cells, as proved by proliferation analyses. Finally, in search for

intrinsic regulators of astroglial types, we found that the abrogation of the transcription

factor Sox2 appears to specifically impact on BG differentiation. In conclusion, this study

demonstrates that cerebellar astrogliogenesis occurs according to a well-defined

spatiotemporal pattern from distinct embryonic and postnatal progenitors, whose fate

potential undergoes a progressive restriction.


44

P4

Zfp423, a Joubert syndrome gene, is a domain-specific regulator of cell cycle

progression, DNA damage response and Purkinje cell development in the cerebellar

primordium

Casoni F. 1,2, Croci L. 1, D’Ambrosio R. 1, Bosone C. 1,2, Sarna J. R5, Warming S. 4.§,

Hawkes R. 5, Consalez G. G. 1,2

The Zfp423 gene encodes a 30-Zn-finger transcription factor (TF) involved in some

regulatory cascades of relevance in cerebellar development. While Zfp423 null mutants

show a significant decrease in the total number of cerebellar Purkinje cells (PCs), the

underlying mechanism remains unclear. Mutations of the human ortholog ZNF423 have

been identified in patients carrying cerebellar vermis hypoplasia or Joubert Syndrome

(JS), associated with other classical ciliopathy signs. ZNF423 also plays a role in the DNA

damage response (DDR). To further characterize the role of ZFP423 in PC development,

we have analyzed two mouse lines carrying allelic deletions of ZFP423. One of them

lacks Zn-finger domain 9-20 (Δ9-20), which mediates functional interactions with BMP

and Notch signaling pathways, and with the DNA repair cofactor PARP1. The other

mutant lacks a C-terminal domain (Δ28-30), which binds to EBF TFs, involved in neuronal

differentiation. In both lines the cerebellar ventricular zone (VZ) features a delay in

progenitor cell cycle progression and an increase in the number of phosphorylated H2A

histone family member X (γH2AX)-positive progenitors, revealing an excess of DNA

breaks in cerebellar VZ progenitors. However, other defects are allele specific. Zfp423

Δ9-20/Δ9-20 mutants exhibit a premature decline of the OLIG2+ PC progenitor pool in

the VZ. In these mutants, M-phase progenitors of the cerebellar VZ display changes in

spindle orientation indicative of a precocious switch from symmetric to asymmetric cell

divisions. Conversely, the Zfp423 Δ28-30/Δ28-30 primordium features a sharp decrease

in the expression of PC differentiation markers, including CORL2. Our in vivo evidence

sheds light on the global and domain-specific roles played by ZFP423 in different aspects

of PC progenitor development, and at the same time supports the emerging notion that

an impairment of the DNA damage response may be a key factor in the pathogenesis of

JS and other ciliopathies.


45

P5

Stereological analysis of Purkinje cell synapse in the molecular layer of the rat

cerebellum according to its phylogenic lobules

Seung Hak Oh, Hyun Wook Kim, Im Joo Rhyu

The cerebellum is a region of the brain that plays an important role in motor control. It is

classified phylogenetically into archicerebellum, paledcerebellum and neocerebellum.

The Purkinje cell is one of the key cells lined in a row, called Purkinje cell layer and it

have a unique dendritic branches with numerous spines.

The previous study reported that there is a difference of synapse density according to the

lobules based on large two-dimensional data. But, recent preliminary data showed there

was no difference in dendritic spine density of the Purkinje cell according to its phylogenic

lobule classification. We designed a stereological analysis of the Purkinje cells synapse

in the molecular layer according to their phylogenic location as a first step to understand

this question.

The 6 weeks old Sparague Dawley rats were perfused and cerebella were dissected and

embedded in resin. We analyzed soma size of the Purkinje cells, their density in the

lobules of II, VI and X with stereological modules such as nucleator and physical

fractionator. The synaptic density was estimated by double disector based on Purkinje

cell density and Purkinje cell synapse density in the molecular layer of the each cerebellar

lobule.

The results showed that there are significant difference in the density of Purkinje cells

and number of synapse per Purkinje cell according to their phylogenic lobules. The

number of Purkinje cell in a given volume was larger in the archicerebellum, but density

of synapse per a Purkinje cell was higher in the neocerebellum.

This data suggest that cellular and synaptic organization of the Purkinje cell is different

according to their phylogenic classification. Further detailed analyses of the dendritic tree

would be important to explain this differential organization.


46

P6

In Vivo 4D MRI of the Developing Mouse Cerebellum

Turnbull DH1, Holmes H1, Rallapalli H1, Suero-Abreu G1, Szulc K1, Tan I2, Joyner AL2

1.Skirball Institute of Biomolecular Medicine, NYU School of Medicine, New York, USA 2.Developmental Biology Program, Sloan Kettering Institute, New York, USA

The early postnatal mouse cerebellum poses a unique challenge for in vivo

developmental imaging studies, with rapidly changing cellular and morphological features

that are difficult to detect and characterize with conventional approaches. High field (≥ 7

Tesla) magnetic resonance imaging (MRI) can be utilized effectively for adult mouse

neuroimaging, but conventional MRI contrast depends on differences in tissue properties

that are largely absent in the developing brain. We have developed 4D (3D + time)

Manganese (Mn)-Enhanced MRI (MEMRI) for in vivo longitudinal analysis of the

developing mouse brain, from fetal stages through the critical neonatal stages of

cerebellar growth and foliation. Non-toxic levels of paramagnetic Mn2+ ions are

introduced by maternal intraperitoneal (IP) injection, and delivered to the pups

noninvasively via lactation. Recent ultra-high resolution images demonstrate that Mn-

uptake and contrast enhancement in the cerebellum is localized to the Purkinje cell layer

and the cerebellar nuclei (CN), allowing exquisite visualization and volumetric analyses

of the developing lobules, and an effective in vivo phenotyping approach for mouse

mutants with defects in CN morphology and cerebellar foliation. The ability to visualize

motor nuclei has also led to applications of MEMRI for in vivo mapping of functional

cerebellar circuits. In addition to imaging cerebellum foliation and nuclei, MEMRI also

provides a sensitive method to detect early pre-neoplastic lesions and to quantify tumor

formation and progression in mouse models of medulloblastoma. These in vivo imaging

methods are providing a quantitative framework for understanding the morphogenesis of

the normal mouse cerebellum, and for analyzing mutant phenotypes and disease in a

wide range of mouse models of cerebellar disorders.


47

P7

Disrupted rhombic lip development caused by aberrant mesenchymal signaling likely

represents a unifying developmental mechanism for human Dandy-Walker malformation

Haldipur P. 1, Dang D. 1, Aldinger KS. 1, Guimiot F. 2, Adle-Biasette H. 2, Bernardo S. 3,

Manganaro L. 3, Silvestri E. 4, Kidron D. 5, Dobyns WB. 1, 6, and Millen KJ. 1, 6

1Center for Integrative Brain Research, Seattle Children's Research Institute, Seattle,

United States 2Hôpital Robert-Debré, INSERM UMR 1141, Paris, France 3Department of Radiological, Oncological and Pathological Sciences, Sapienza

University of Rome Policlinico Umberto I Hospital, Rome, Italy. 4Surgical Pathology Unit, San Camillo Forlanini Hospital, Rome, Italy. 5The Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel. 6Department of

Pediatrics, Genetics Division, University of Washington, Seattle, United States

Human cerebellar malformations are recognized with relative ease through brain imaging

studies. However, the molecular and cellular mechanisms contributing to cerebellar birth

defects are poorly understood and their developmental pathology is largely undescribed.

We have reported that in rare patients, FOXC1 loss contributes to a posterior fossa

phenotypic spectrum that includes Dandy-Walker malformation (DWM), a common

human cerebellar malformation. We now demonstrate that the null and hypomorphic

Foxc1 mutant mice have early granule and Purkinje cell (PC) abnormalities and

subsequent disruptions in cerebellar foliation and lamination. Particularly striking is the

presence of a partially formed unpaired posterior vermis lobule which echoes the

posterior vermis DW “tail sign” observed in human imaging studies. Lineage tracing

experiments in both the null and hypomorphic Foxc1 mouse mutants indicate that the

main cause of this feature is the aberrant migration of granule cell progenitors from the

rhombic lip that are destined to form the posterior-most lobule. This phenotype is due to

loss of required signaling molecules including SDF1 from the mesenchyme surrounding

the developing cerebellum. Analyses of rare human DW fetal cerebella with chr 6p25

(FOXC1) heterozygous deletions demonstrate extensive phenotypic overlap with our

Foxc1 mutant mouse models, validating our DWM models and demonstrating that many

key mechanisms controlling cerebellar development are conserved between mouse and

human. Ongoing analysis of additional DWM fetal samples of unknown genotypes

demonstrates remarkably similar features, suggesting that we have identified a unifying

developmental mechanism for DWM.


48

P8

Identification of key regulators for cerebellar development using FANTOM5 time-course

CAGE data

Thomas J. Ha1*, Anthony Mathelier1, 2, 3*, Peter Zhang1*, Remi Robert1, Tyler Funnel4,5,

The FANTOM Consortium, Wyeth W. Wasserman1, Daniel Goldowitz1**

1 Centre for Molecular Medicine and Therapeutics at the Child and Family Research

Institute, Department of Medical Genetics, University of British Columbia, Vancouver,

BC, Canada, 2Centre for Molecular Medicine Norway (NCMM), Nordic EMBL

Partnership, University of Oslo, 0318 Oslo, Norway, 3Department of Cancer Genetics,

Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, 0372 Oslo,

Norway, 4Department of Molecular Oncology, BC Cancer Agency, 675 W10th Avenue,

Vancouver, BC, V5Z 1L3, Canada, 5Bioinformatics Graduate Program, University of

British Columbia, Vancouver V5Z 1L3, BC, Canada *These authors contributed equally to this work ** Corresponding Author: [email protected]

The work of the FANTOM5 Consortium has brought forth a new level of understanding

of the promoter regulation and cellular processes involved in creating diversity of cell

types. In this study, we extended the analysis of the FANTOM5 Cap Analysis of Gene

Expression (CAGE) transcriptome data to focus on understanding the genetic regulatory

mechanisms involved in mouse cerebellar development. We performed

HeliScopeCAGE library sequencing on cerebellar samples over 8 embryonic and 4

early postnatal times. This study showcases temporal expression pattern changes

during cerebellar development. We have completed a bioinformatics analysis that

focused on the transcription factors, their promoters and binding sites which identifies

genes that appear as strong candidates for involvement in cerebellar development. We

selected several candidate cerebellar gene regulators for validation experiments

including qRT-PCR, immunocytochemistry and shRNA transcript knockdown. We

observed severe developmental defect in Atf4, Rfx3 and Scrt2 knockdown embryos,

which indicate these three genes as key regulatory genes in cerebellar development.

More importantly, the successful identification of these novel cerebellar gene regulator

demonstrated that the FANTOM5 cerebellum time series is an accessible, high-quality

transcriptome database for functional investigation of gene regulatory networks in

cerebellar development.



49


Theme 2- Cerebellar Normal and Abnormal Differentiation.

P9 / Miss Xiaodan Jiao/ The sonic hedgehog signaling pathway in development of cerebellar

granule cells

P10 / Miss Niloufar Ashtari/ Cerebellar corticogenesis in the lysosomal acid phosphatase (Acp2)

mutant mouse: Purkinje cell migration disorder

P11 / Miss. Margaret Stromecki/ OTX2 controls an axon guidance gene expression network to

regulate medulloblastoma self-renewal.

P12 / Miss. Lisa Liang/ CD271 (p75 Neurotrophin Receptor) as a novel diagnostic marker and

therapeutic target in sonic hedgehog medulloblastoma.

P13 /Prof. Jerry Vriend/ Differential expression of genes for proteasome subunits and ubiquitin

ligases in medulloblastoma subtypes.


50

P9

The sonic hedgehog signaling pathway in development of cerebellar granule cells

Jiao X, Ashtari N, Rahimi Balaei M, and Marzban H

Department of Human Anatomy and Cell Science, The Children’s Hospital Research



[email protected]

Introduction: During development, cerebellar granule cell precursors arise from the

rhombic lip and form the external germinal zone. The granule cell precursors proliferate

and then migrate to the granule cell layer and differentiate into mature granule cells. In

this process, the sonic hedgehog (Shh) and N-myc pathways are important promoters for

granule cell precursor proliferation. The lysosomal acid phosphatase 2 (Acp2) mutant

mouse (naked-ataxia, nax) shows a significant reduction in granule cells in the

cerebellum. We hypothesize that the decrease N-myc expression interrupts the Shh

pathway in nax cerebellar granule cells development.

Methods: We used nax mice and wild-type siblings as controls in this study. In vivo and

in vitro immunohistochemistry and Western-blotting were used to detect molecular

expression.

Results: In the nax mouse, there is an 80% reduction in cerebellar granular cells

compared with wild-type sibling mice during postnatal development. The Shh expression

changed with a delay in the nax cerebellum compared with wild-type. Our data showed

strikingly reduced N-Myc expression in the nax cerebellum, which was accompanied by

an increase in proteasome activity.

Conclusion: This study suggests that Shh and the N-myc pathway are impaired in the nax

mouse and granule cells proliferation is prevented during cerebellar development. The

significant reduction in proliferation and probably differentiation of granule cells in the nax

mouse reveals that Acp2 mutation affects proteasome activity and dysregulates N-myc

expression in the Shh pathway.


51

P10

Cerebellar corticogenesis in the lysosomal acid phosphatase (Acp2) mutant mouse:

Purkinje cell migration disorder

Ashtari N, Jiao X, Rahimi Balaei M, Ghavami S, and Marzban H

Department of Human Anatomy and Cell Science, The Children’s Hospital Research



[email protected]

Introduction: A mouse mutant called nax (naked-ataxia), resulting from a spontaneous

mutation in the lysosomal acid phosphatase (Acp2) gene, shows severe cerebellar

defects and neuronal developmental disorders in its cerebellum. In the Acp2 mutant

mouse, three layers of the cortex were significantly impaired and monolayer Purkinje cells

(Pcs) turn into multi-layered Pcs that ectopically invade the molecular layer, indicating

that they have no stop signal. We investigated the Reelin-VLDLr-Dab1 signaling pathway,

which is important in Pcs migration and monolayer formation in the cerebellar cortex.

ERK1/2, which is a member of the mitogen activated kinase family and downstream of

Reelin signaling, was examined. We hypothesize that the establishment of mono-layered

Pcs is dependent on Reelin through the ERK1/2 signaling pathway.

Methods: Acp2 mutant mice were used for this study and molecular expression and

distribution were assessed by immunohistochemistry, Western blotting, and cell culture.

Results: The cerebellar cortex of Acp2 mutant mice revealed the presence of Pcs in a

randomized and dispersed manner spanning the entire molecular and Pc layers, rather

than a monolayer in the cerebellar cortex. The pattern of Reelin expression showed a

down-regulation in both wild type and nax mice, while less protein is detected in the nax

mutant (significantly at P4, P7 that is around Pc layer formation) compared with the wild

type during postnatal development. QPCR examination showed that other molecules

downstream of Reelin including VLDLr and Dab1 have a higher transcript expression level

in nax. In addition, the ERK1/2 expression level is increased in the nax mutant mouse.

Conclusion: These results advance our understanding of the mechanism of Pcs

migration. These observations suggest that less Reelin expression may lead to increased

VLDLr and Dab1 levels in the nax mutant mouse cerebellum. Because Reelin can

modulate expression of the ERK1/2 pathway, our data suggest that overexpression of

ERK1/2 in nax mice may affect the Pcs stop signal to establish their proper position and

form a multilayer of Pcs during cerebellar development.



52

P11

OTX2 controls an axon guidance gene expression network to regulate medulloblastoma

self-renewal

Stromecki M. 1, Tatari N. 1, Morrison L. 1, Kaur R. 1, Palidwor G. 2, Porter C. 2, Skowron

P. 4 , Wölfl M. 3, Taylor M. 4, Werbowetski-Ogilvie T. 1

1 University of Manitoba, Department of Biochemistry and Medical Genetics, Winnipeg,

Canada; 2 Ottawa Hospital Research Institute, Ottawa, Canada; 3 University of Wuerzburg, Würzburg, Germany; 4 The Hospital for Sick Children, Arthur and Sonia Labatt Brain Tumour Research

Centre and Program in Developmental and Stem Cell Biology, Toronto, Canada.

Medulloblastoma (MB) is a highly heterogeneous primary pediatric brain cancer that is

frequently accompanied by metastatic dissemination and poor long-term prognosis. The

most aggressive tumors are refractory to conventional chemotherapy and radiation. Our

goal is to identify new signaling pathways that regulate the treatment-resistant MB cellular

phenotypes, namely the stem cells. We previously discovered that the OTX2 gene, which

is amplified or overexpressed in the majority of aggressive MBs, is a central regulator of

stem cell function or self-renewal in these tumors. However, the molecular mechanisms

by which OTX2 regulates these functions are still unknown. Here, we employed

complementary bioinformatics approaches to characterize the OTX2 regulatory network

and identified a novel relationship between OTX2 and genes associated with axon

guidance signaling in Group 3 and Group 4 MB stem/progenitor cells. ChIP-sequencing

identified putative OTX2 DNA binding sites and revealed statistically significant

associations between OTX2 and these axon guidance pathway genes. Group 3 and 4

MB patient samples were evaluated for correlations between expression of axon

guidance pathway genes, OTX2 and survival. Semaphorin signaling was the most

overrepresented pathway across all datasets with expression of all pathway genes being

upregulated following OTX2 KD. Moreover, SEMA4D expression was identified as a novel

prognostic biomarker. Functional validation studies demonstrated that increased levels of

semaphorin pathway genes are associated with a decrease in self-renewal underscoring

a potential tumor suppressive role in MB. Downstream pathways known to mediate the

effects of semaphorin signaling including, RHOA and MAPK were differentially expressed

following OTX2 KD. Our results offer critical insights into the molecular drivers of the most

aggressive MB tumors and provide an informed framework to pursue novel targeted

therapies aimed at differentiating MB tumors cells.


53

P12

CD271 (p75 Neurotrophin Receptor) as a novel diagnostic marker and therapeutic

target in sonic hedgehog medulloblastoma

Liang, L 1, Coudiere-Morrison, L 1, Tatari, N 1, Ramaswamy, V 2, Ryken, T 4, Del Bigio,

D 5, Taylor, M 2, Hawkins, C 2, Chan, J 6, Werbowetski-Ogilvie, T 1

1 Regenerative Medicine Program, Department of Biochemistry and Medical Genetics,

University of Manitoba, Winnipeg, Manitoba, Canada 2 Arthur and Sonia Labatt Brain

Tumour Research Centre and Program in Developmental and Stem Cell Biology, The

Hospital for Sick Children, Toronto, Ontario, Canada 3 Cancer Care Manitoba (CCMB),

Winnipeg, Manitoba, Canada 4 Department of Neurosurgery, University of Kansas,

Kansas City, Kansas 5 Department of Pathology, University of Manitoba and Manitoba

Institute of Child Health, Winnipeg, Manitoba, Canada 6 Department of Pathology and

Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada

Medulloblastoma (MB) is the most common malignant primary brain tumor and is

currently classified into 5 distinct molecular subtypes. This extensive heterogeneity has

revealed a critical need for subtype-specific, functionally validated biomarkers and

therapeutic strategies. Using a high throughput flow cytometry screen and gain/loss of

function studies, we previously identified CD271/p75NTR as a candidate stem cell

marker, specifically in SHH MB. Here, we show that CD271+ and CD271- subpopulations

do not exhibit a hierarchal organization; but are rather two co-existing cellular subsets.

FACS sorting followed by re-culturing of CD271- and CD271+ cells from low passage

primary cultures demonstrated that both cell populations can recapitulate the parental

phenotype. However, global gene expression profiling by RNA sequencing revealed that

genes/pathways associated with cell survival, proliferation and motility are downregulated

in CD271- vs. CD271+ subpopulations. Cell cycle analysis supported these findings and

demonstrated that CD271- cells exhibited a decrease in proliferation compared to

CD271+ cells. Moreover, PI3K/AKT, JAK/STAT, and NFkB, are downregulated in CD271-

vs. CD271+ cells, while pathways such as RAS/MYC are upregulated in CD271- vs.

CD271+ cells. IHC analysis of CD271 levels across the MB subtypes demonstrated that

nearly all SHH tumors express CD271, while Group 3 tumors exhibit moderate staining.

In contrast, Group 4 MB and WNT do not express CD271. Importantly SHH MB exhibit a

nodular CD271 staining pattern, further underscoring the heterogeneity within these

tumors and supporting our findings that CD271- and CD271+ cells are distinct, yet

clinically relevant, subpopulations. Our results suggest that CD271, in combination with

other markers, could be effectively utilized as a diagnostic tool for SHH MB and that

therapeutic strategies concomitantly targeting both CD271+ and CD271- subpopulations

would be the most effective in SHH MB treatment.


54

P13

Differential expression of genes for proteasome subunits and ubiquitin ligases in

medulloblastoma subtypes

Jerry Vriend

Department of Human Anatomy and Cell Science, University of Manitoba

Introduction; A study of gene expression in a group of 46 medulloblastomas (MB) reported

differential gene expression of more than 2600 genes among five different subtypes of

MB in 2006 (The Thompson dataset). The Thompson dataset later contributed to a

consensus classification in which there are four MB molecular subtypes (Wnt, SHH,

Group 3 and Group 4). Investigators of the Thompson report made available data in

which they tabulated the expression of many genes not discussed, including those coding

for proteasome subunits and ubiquitin ligases. Several additional available datasets

supporting the consensus classification also included data on expression of genes for

proteasome subunits and for ubiquitin ligases.

Methods and Objectives; Using the Ubiquitin and Ubiquitin-like Conjugation Database

(UUCA) we screened the various datasets for the purpose of determining whether

expression of genes of the ubiquitin proteasome pathway were differentially expressed

among the subtypes of MB identified in these studies. We then compared gene

expression in the Thompson dataset to that of other publically datasets of

medulloblastoma subtypes.

Results and Conclusions; Genes with significant differential expression among MB

subtypes included genes for core proteasome subunits, genes for ubiquitin conjugating

enzymes, genes for numerous ubiquitin E3 ligases and genes for several deubiquitinases.

The Thompson dataset thus provides evidence that the expression of genes for core

proteasome subunits, as well as the genes for numerous ubiquitin ligases, contribute to

the classification of medulloblastomas into subtypes. Much of the Thompson data was

confirmed by datasets of Kool, Gilbertson, Pfister and Northcott. The data show that

genes for components of the ubiquitin proteasome system contribute to the molecular

classification of MBs. They should be investigated further as therapeutic targets.


55


Theme 3- Cerebellum Circuitry and Functional development.

P14 / Dr. James M. Bower/ The ascending branch of the granule cell axon: Implications for

development and function.

P15 / Miss. Julie Marocha/ Clustered protocadherin diversity in Purkinje cell dendrite

development.

P16 / Prof. Kin Ming Kwan/ Lhx1/5 is critical in controlling dendritogenesis and spine

morphogenesis of Purkinje cells via regulation of Espin.

P17 / Miss. Wendy Wang/ Regulation of Cerebellar Interneuron Morphology and Integration by

the Clustered Protocadherins.

P18 / Dr. Mehdi Mehdizadeh/ Dopamine Receptors Expression in Lysosomal Acid Phosphatase

(Acp2) Mutant Mice Cerebellar Cortex

P19 / Mr. Zachary Nurcombe/ The expression of the small heat shock protein Hsp25 in the

developing rodent cerebellum.

P20 / Miss. Jennifer Flood/ The expression of the small heat shock protein Hsp25 in the

embryonic chick cerebellum.


56

P14

The ascending branch of the granule cell axon: Implications for development and

function.

Bower, James M. 1

1Virtual Worlds IP Inc.

Almost 40 years ago, as a first year graduate student newly joining the laboratory of Dr.

Wally Welker at the University of Wisconsin, I had an idea for an experiment. Using high-

density micro-electrode mapping techniques, Dr. Welker and his colleagues had recently

discovered an unusual ‘fractured’ pattern of tactile afferent projections to the granule cell

layers of the lateral hemispheres of the rat cerebellum. The maps were of a finer grain

than any previously recorded and immediately suggested that overlying Purkinje cells

should respond, via the parallel fibers, to tactile inputs originating from many locations on

the body surface. Accordingly, I proposed to explore parallel fiber driven Purkinje cell

activity in response to the multiple different types of tactile inputs projecting to the granule

cell layer. Surprisingly, however, the recorded Purkinje cell responses suggested that

parallel fibers did not directly drive Purkinje cell output. Those results and 40 years of

subsequent experimental and modeling results suggest instead that the granule cells

provide two distinctly different types of synaptic influences on Purkinje cells with the

primary influence on somatic spiking provided by synapses associated with the ascending

segment of the granule cell layer, modulated by a much more subtle influence of the

parallel fibers. These results in turn suggest that the cerebellar cortex has a very different

functional organization than assumed by most theories of its function. This poster will

first provide evidence for this dual granule cell influence and then consider the

implications for both the development of the cerebellar cortex as well as its function and

dysfunction.


57

P15

Clustered protocadherin diversity in Purkinje cell dendrite development

Marocha J. 1, 2, Lefebvre J.L. 1, 2

1 The Hospital for Sick Children, Program for Neuroscience and Mental Health, Toronto,

Canada 2 University of Toronto, Department of Molecular Genetics, Toronto, Canada

Dendrites of individual neurons develop non-overlapping arbors through a mechanism

called neurite self-avoidance. By evenly spacing out their dendrites, neurons maximize

their receptive fields for effective sampling of inputs and minimize the likelihood of making

self-connections. In some cases, neurons that exhibit self-avoidance also interact

extensively with dendrites of neighbouring neurons of the same type suggesting that

neurons can discriminate 'self' dendrites from 'non-self'. The clustered Protocadherins

(Pcdhs), a large group of cell surface molecules, have been implicated in these

processes, as they provide enormous molecular diversity and single neuron identity. The

Pcdh locus is divided into the alpha (α), beta (β), and gamma (γ) gene clusters and

encodes 58 isoforms. Pcdh isoforms exhibit homophilic recognition properties and have

been shown to be randomly and combinatorially expressed among individual neurons.

We have previously shown that γ-Pcdhs mediate dendrite self-avoidance in cerebellar

Purkinje cells (PCs) (Lefebvre et al, Nature 2015). The first objective of this study was to

determine whether α-Pcdhs also signal in PC dendrite self-avoidance. The second

objective of this study was to investigate the functional interactions between members of

multiple Pcdh clusters. Behavioural outcomes of deleting Pcdhs from cerebellar cells in

mice will be discussed. I will also present ongoing work elucidating the cellular and

homotypic mechanisms underlying dendrite self-avoidance. One major finding in my work

was that α-Pcdhs are also required for dendrite self-avoidance. I used an adenovirus

expressing GFP to selectively label and analyze individual PCs. This study will contribute

to our understanding of the molecular mechanisms of PC dendrite development and the

impact of dendritic patterning on cerebellar-related functions.


58

P16

Lhx1/5 is critical in controlling dendritogenesis and spine morphogenesis of Purkinje

cells via regulation of Espin

Kwan K.M. 1, 2, 3, Lui N.C. 1, Tam W.Y. 1, Gao C. 1, Wang C.C. 4, 5, 6, Jiang L. 1, 2, 3, Yung

W.H. 5, 7, Huang J. 8

1 School of Life Sciences, 2 Center for Cell & Developmental Biology, 3 Partner State

Key Laboratory of Agrobiotechnology (CUHK), 4 Department of Obstetrics and

Gynecology, 5 School of Biomedical Sciences, 6 Li Ka Shing Institute of Health

Sciences, 7 Gerald Choa Neuroscience Centre, The Chinese University of Hong Kong,

Shatin, Hong Kong, China. 8 Department of Biochemistry, University of Hong Kong,

Pokfulam, Hong Kong, China.

Purkinje cells (PCs), the only output neurons in cerebellar cortex, have extensively

branched dendrites to receive signals from different cerebellar neurons and serve as an

integration centre in cerebellar cortex. Defects in the dendritic development of PCs thus

disrupt cerebellar circuitry and cause ataxia. However, the molecular mechanism of

dendritic development remains unclear. Our group found that the specific inactivation of

both Lhx1 and Lhx5 in postnatal PCs resulted in ataxic mutant mice with abnormal PC

dendritic development. The PCs of Lhx1/5 mutants had reduced expression of Espin, a

novel F-actin cytoskeleton regulator. We later identified that Espin expression was

transcriptionally activated by Lhx1/5. Downregulation of Espin in the PCs caused F-actin

mislocalization and impaired dendritogenesis and spine morphogenesis. The mutant PCs

could not properly innervate with the pre-synaptic inputs, leading to aberrant

electrophysiological properties. By overexpressing Espin in the mutant PCs, we were able

to rescue the F-actin localization defects and the dendritic defects in the mutant PCs. Our

findings give evidences for a novel pathway controlling dendritic development in which

Lhx1/5, through regulating Espin expression, govern dendritogenesis and spine

morphogenesis in postnatal PCs.


59

P17

Regulation of Cerebellar Interneuron Morphology and Integration by the Clustered

Protocadherins

Wang W. 1, 2, Lefebvre J.L. 1, 2

1 Program for Neuroscience and Mental Health, the Hospital for Sick Children, Toronto,

Canada 2 Department of Molecular Genetics, University of Toronto, Toronto, Canada

The large diversity of neurons can be subdivided into morphological classes with distinct

connectivity patterns, suggesting that cell-type specific morphologies are critical for

function. The cerebellar cortex, with its protracted timeline of development and

stereotyped cellular architecture, is an ideal model to study mechanisms that shape

neuronal arborization and circuit assembly. Improper development or maintenance of this

circuitry is implicated in several neurodevelopmental and neurological disorders, such as

autism and spinocerebellar ataxias. Proper development of the cerebellar circuitry

requires coordinated and cell-type specific actions of recognition proteins, but molecules

and mechanisms that contribute to precise wiring are poorly understood. Here, I present

my work on the roles of the clustered Protocadherin (Pcdh) family of cell-surface

receptors in neurite arborization of the Molecular Layer Interneurons (MLIs). With a

remarkable potential for molecular diversity and wiring specificity, Pcdhs are expressed

in multiple cell-types in the cerebellum. We have previously shown that Pcdhs regulate

Purkinje cell dendrite arborization. To investigate how Pcdhs pattern other cerebellar cell-

types, and whether they regulate circuit assembly through cell-cell interactions, I

interrogated the consequences of Pcdh deletion in MLIs at various stages of integration.

Using conditional mutant mouse alleles and techniques to sparsely label MLIs, I show

that loss of the gamma-Pcdh isoforms (Pcdh-γ) lead to decreased MLI dendritic

branching, confirming cerebellum-wide roles in dendrite arborization. In addition, MLIs in

Pcdh-γ mutants have decreased axonal innervations, suggesting novel roles in axonal

patterning. These findings implicate the Pcdhs in regulating neurite patterning of

cerebellar MLIs. My future work aims to delineate the roles of Pcdh-dependent isoform

diversity and cell-cell interactions in proper establishment of MLIs within the cerebellar

circuitry.


60

P18

Dopamine Receptors Expression in Lysosomal Acid Phosphatase (Acp2) Mutant Mice

Cerebellar Cortex

Mehdizadeh M.1, Ashtari N.2, Rahimi Balaei M.2, Jiao X.2, and Marzban H.2

1Department of Human Anatomy, Iran University of Medical Sciences, Tehran, Iran 2Department of Human Anatomy and Cell Science, The Children’s Hospital Research


science, University of Manitoba, Winnipeg, MB, R3E 0J9, Canada.

[email protected]

Introduction: In mice, a spontaneous mutation in Lysosomal Acid Phosphatase (ACP2)

enzyme results in severe cerebellar defects. These include a reduced size, abnormal

lobulation, and neuronal migration disorders with an absent or hypoplastic anterior

vermis. Dopamine receptors are involved in cerebellar development and functions. In

order to better understand the specific interactions of dopaminergic system in developing

cerebellum, we studied the dopamine receptors expression in the cerebellum of the nax

mutant mouse.

Methods: Anti-dopamine receptors antibodies (D1, D2, D3, D4, and D5) were used to

perform immunohistochemistry from P1 to P18 in nax mutant and wild type sibling

cerebellar tissues.

Results: Immunohistochemistry revealed D1 and D2 receptor expression in Purkinje cells

and cerebellar nuclei neurons and was increased in nax cerebellum in comparison to the

wild type sibling. There were no differences in D3 receptor expression and were mainly

expressed in Purkinje cells, cerebellar nuclei neurons and molecular layer interneurons.

The D4 receptor expression was patterned in Purkinje cell and molecular layers, while D5

receptor expression was mostly localized in nerve fibers with slight differences between

nax and wild type sibling cerebellum.

Conclusion: This study contributes to our understanding of the distribution of the

dopaminergic system in cerebellum. Our result shows alteration in the expression pattern

of dopamine receptors in nax mutant cerebellum in comparison to the wild type sibling.

These observations highlight the role of dopaminergic innervation in the cerebellum and

suggest the role of Acp2 in regulating the dopaminergic system.


61

P19

The expression of the small heat shock protein Hsp25 in the developing rodent

cerebellum.

Nurcombe Z 1, Gorrie J 2, Flood JM 1, Armstrong CL 1, 3

1Mt Royal University, Calgary AB CANADA 2University of Guelph, Guelph ON CANADA 3University of Calgary, Calgary AB CANADA

The small heat shock protein Hsp25 is expressed in stripes of Purkinje cells in the central

and nodular zones of the adult mouse cerebellum. This expression is constant in

numerous strains of mice (e.g. CD1, BalbC, C3H, C57Bl6, etc) and not altered by

perturbation (e.g heat shock, afferent fiber ablation etc). In contrast, Hsp25 is not seen

in the adult cerebellum of the KJR/MsKist mouse line (originally derived from Korean wild

mice) or in the adult rat cerebellum. In an attempt to understand this variability and the

role of Hsp25 in cerebellar Purkinje cells, we used western blot analysis and

immunohistochemistry to look at the developmental expression of Hsp25 in the postnatal

KJR and rat cerebella and compare it to that seen in the developing wildtype mouse

cerebellum.

Hsp25 expression in the developing wildtype mouse cerebellum is dynamic in that Hsp25

is initially turned on in a few clusters of Purkinje cells shortly after birth, and then widely

expressed in the majority of Purkinje cells, and then refined into the stripe pattern seen in

the adult.

Preliminary results of this study show that despite the absence of Hsp25 in the KJR adult

cerebellum, there is a similar pattern of expression seen during KJR development albeit

in a compressed time frame. In the developing KJR cerebellum, Hsp25 was widely

expressed in Purkinje cells at postnatal day (P) 4, refined into stripes by P8 and then gone

by P12. In contrast, aside from a few scattered Hsp25-immuopositive Purkinje cells seen

in the anterior zone of the neonatal rat cerebellum, Hsp25 was not seen in either the

developing or adult rat cerebellum.


62

P20

The expression of the small heat shock protein Hsp25 in the embryonic chick

cerebellum.

Flood JM 1. Lim D 2, Gilbert EA 2, Kameka A 3, Armstrong CL, 1, 3

1Mt Royal University, Calgary AB CANADA 2University of Guelph, Guelph ON CANADA 3University of Calgary, Calgary AB CANADA

The embryonic chick is an ideal model for studying pattern formation in the cerebellum

due to the easily accessible embryo at all stages of development. The newly-hatched

chick is precocial - able to stand, walk and run within hours. Despite a similar length

gestation, mouse pups are altricial (immature, immobile) at birth and locomotor

development takes weeks: it is plausible that this difference in coordinated motor

movement may be reflected in the timing of protein expression patterns in the cerebellum.

Previous findings in our lab have demonstrated that Zebrin II, a ‘patterning’ antigen

expressed in a specific subset of Purkinje cells is expressed in posterior folia as early as

embryonic day 12 (compared to ~postnatal day 5 in the mouse cerebellum). ZII

expression is robust by E14-E16 and ZII-immunopositive and -negative stripes of Purkinje

cells emerge at E18-20 (Gilbert et al., 2012).

In the present study, we used western blot analysis and immunohistochemistry to

determine the expression of an additional Purkinje cell ‘patterning’ antigen - the small

heat shock protein Hsp25. Preliminary results suggest that, like ZII, Hsp25 is initially

expressed between E12-E14. From E16 through to hatch, Hsp25 is widely expressed in

Purkinje cells throughout folia of embryonic chick cerebellum. This embryonic expression

precedes that seen in neonatal mice by nearly a week. The earlier/accelerated timing of

these antigens in the embryonic chick cerebellum as compared to the mouse may reflect

the precocial character (e.g. motor co-ordination, mobility) of chicks at hatch. This study

will broaden our understanding of the specific patterning and organization of the

developing avian cerebellum and provide the foundation for further studies designed to

investigate the function of Hsp25 in cerebellar Purkinje cells.


63


Theme 4- Aberrations of Cerebellar Development and Function: Genetics and Imaging

P21 / Miss. Suteera Vibulyaseck/ Rearrangement of domains of protocadherin 10-positive

Purkinje cell subsets in the mouse embryonic cerebellum.

P22 / Dr. Ryan Willett/ The cerebellar nuclei dictate the size of the cerebellum by regulating

Purkinje cell number.

P23 /Dr. Xavier Guell Paradis/ Functional topography of the human cerebellum: An fMRI study

using the Human Connectome Project dataset.

P24 / Dr. Kimberly Aldinger/ Insights into the genetic landscape of cerebellar malformations.

P25 / Dr. Shervin Pejhan/ Methyl-CPG binding protein 2 isoform-specific regulatory network in

human cerebellum.

P26 / Dr. Jan Cendelin/ Embryonic cerebellar graft development in the cerebellum of normal and

cerebellar mutant mice.


64

P21

Rearrangement of domains of protocadherin 10-positive Purkinje cell subsets in the

mouse embryonic cerebellum

Vibulyaseck S1, Fujita H1, 3, Sugihara I1, 2

1 Tokyo Medical and Dental University, department of Systems Neurophysiology 2 Center for Brain Integration Research, Tokyo, Japan 3 Johns Hopkins University, department of Otolaryngology-HNS, Baltimore, U.S.A.

The mammalian cerebellum is subdivided into several compartments that consist of

transversely-oriented lobules and longitudinally-arranged Purkinje cell (PC) stripes. The

anterior and posterior lobules are associated with somatosensorimotor function while the

central lobules are involved in non-motor function. The PC monolayer reveals

heterogeneous compartments that have different molecular compositions; among these

is Aldolase C (Aldoc) which is a late-onset marker. Therefore, we looked at

Protocadherin10 (Pcdh10) which is expressed before birth, and hence can help us

understand this compartmental pattern. In the late embryonic day (E) 17.5, we have

previously found that clusters are formed already as some 50 Purkinje cell clusters.

However, the relationship between embryonic cluster formation before E17.5 and adult

stripes is not much clarified yet. In this present study, first, PC subsets are reconstructed

at E14.5 to see their spatial distribution in the entire mouse cerebellar cortex. Then the

expression of Pcdh10-positive subareas, visualized in OL-KO knock-in mouse strain, are

shown in some particular domains as defined by medial, dorsal, central, and mid-lateral

groups from E14.5 to E17.5. We tracked the change of these domains in shape and

position by reconstructing them in the three-dimensional space from serial sections. We

found that these domains of PC subsets separated and shifted into particular lobules with

immature fissures separating them. While the longitudinally-shifting domains settled in

the anterior and posterior lobules, transversely-shifting domains settled in the central

lobules. The results indicate that the process of formation of multiple stripes in adult

cerebellar cortex occur during the early stages by the separation and shifting of domains

of PC subsets. Support: KAKENHI 16K070025. No conflict of interest.


65

P22

The cerebellar nuclei dictate the size of the cerebellum by regulating Purkinje cell

number

Willett R.T., Roberts K.J., Tournaire G, and Joyner A.L.

Memorial Sloan Kettering Cancer Center, Developmental Biology Program, New York,

USA

The cerebellar nuclei (CN) are specialized neuronal assemblies - collectively comprised

of the fastigial, interpositus and dentate nuclei - that reside in the confluence of white

matter in the cerebellar interior. The CN operate as signal integration centers and relay

posts between the cerebellar cortex, cerebellar afferents and the targets for their efferents

elsewhere in the brain. With the exception of the posterior vermis, Purkinje cells (PCs)

project to the CN. PCs function as a physiological hub, integrating information from

cerebellar afferents and activity within the cerebellar cortex, as well as a developmental

hub secreting Sonic Hedgehog (SHH) that stimulates proliferation of progenitors of

interneurons and astrocytes, as well as granule cell precursors (GCPs). Cerebellar growth

is largely driven by the SHH-driven expansion of GCP numbers and their concomitant

differentiation. Surprisingly, we found that conditional knockout of the homeobox genes

En1 and En2 (En1/2) in only the CN projection neurons (CN-En1/2) results in greatly

reduced cerebellar growth with preferential disruption of particular lobules. In contrast,

loss of En1/2 in GCPs does not cause hypoplasia. We demonstrate that in CN-En1/2

mutants, CN development proceeds normally, but as the projection neurons descend into

the cerebellum, the medial Tbr1+ nuclei cells become disordered and then some cells are

lost between E15.5-E16.5. This CN cell loss is accompanied by a cell non-autonomous

decrease in the number Purkinje cells (PCs), and a delay in their maturation and reduced

Shh expression. Strikingly, throughout development, the vermis hypoplasia phenotype of

CN-En1/2 mutants is accompanied by a normal cerebellar cortical cytoarchitecture, as

the arrangement and proportions of cell types are preserved. Thus, the number of

neurons in the cerebellar cortex scales to match their earlier born output neurons by fine-

tuning proliferation and cell viability, and thus ensuring proper circuit design.


66

P23

Functional topography of the human cerebellum: An fMRI study using the Human

Connectome Project dataset.

Guell X. 1, 2, Gabrieli J.D.E. 1, Schmahmann J.D. 2

1 Massachusetts Institute of Technology, Cambridge, USA; 2 Massachusettes General Hospital and Harvard Medical School, Boston, USA.

Evidence suggests that the cerebellum is engaged in cognition and affect as well as motor

control; however, its contribution to such functions is incompletely understood. One

central question is whether functional subregions exist in the cerebellum and, if so, how

they are organized. The conclusion that the cerebellum is organized into multiple motor

and nonmotor functional subregions is derived from a previous meta-analysis and

prospective study of nine subjects by our group (Stoodley and Schmahmann, 2009;

Stoodley et al., 2010, 2012), and from the meta-analysis of E et al (2014).

We used data of the Human Connectome Project (HCP; n=787) to analyze cerebellar

fMRI task activity (motor, working memory, language, social and emotion processing) and

resting-state functional connectivity. The latter was calculated using cerebral cortical

seeds placed at the peak Cohen’s d of each task contrast.

Our results replicate and extend previous findings of cerebellar functional topography.

We observed motor task activation in lobules IV/V/VI and VIII. In nonmotor tasks,

activation was situated in lobules VI and VII. We did not replicate previous observations

of activation in lobules IV/V and VIII by nonmotor tasks, further supporting the selective

engagement of these lobules in sensorimotor control. Our analysis also revealed

nonmotor activation in lobules IX/X, consistent with some previous observations.

Further, we compared topographical patterns of motor and nonmotor activity as well as

resting-state functional connectivity from the HCP with classical studies using

electrophysiology, tract-tracing, and grey matter morphometry. This was inspired by

current trends in neuroscience suggesting a close relationship between anatomical

connections, task activity, resting-state functional connectivity and pathological grey

matter decrease. Using this approach, we were able to investigate potential overarching

principles of cerebellar organization across multiple topographical domains.


67

P24

Insights into the genetic landscape of cerebellar malformations

Aldinger KA1 Timms AE2 Chung V1 Dobyns WB1, 3

1 Center for Integrative Brain Research, Seattle Children’s Research Institute, Seattle,

USA 2 Center for Developmental Biology and Regenerative Medicine, Seattle Children’s

Research Institute, Seattle, USA 3 Department of Pediatrics, University of Washington, Seattle, USA

Cerebellar malformations are common, heterogeneous birth defects that are associated

with significant developmental disabilities, including coordination problems, epilepsy, and

intellectual and psychiatric impairments. Both genetic and non-genetic prenatal factors

have been implicated in the cause of cerebellar malformations. However, few patients

receive a genetic diagnosis and, for most patients, no underlying cause is ever identified.

Objective: To address the genetic basis of cerebellar malformations, we performed

exome sequencing in 100 families with one or more children diagnosed with Dandy-

Walker malformation (DWM), cerebellar vermis hypoplasia (CBVH), or diffuse cerebellar

hypoplasia (CBLH).

Results: We identified an underlying genetic cause in 40 of 100 families. Among the 49

variants identified, 55% (27/49) were de novo, 70% (19/35) were missense (19/35), and

24% (12/49) were already known to be pathogenic. Our results include 27 genes

previously associated with neurodevelopmental disorders, including 6 genes with

mutations found in more than one patient, and 5 novel genes. Overall, we obtained a

genetic diagnosis for 25% of children diagnosed with classic DWM and for 52% of children

diagnosed with other cerebellar malformations.

Conclusions: Our data show that exome sequencing is a powerful tool for establishing

molecular diagnoses for children diagnosed with common cerebellar malformations. We

also newly implicate several genes in cerebellar development and further show that

cerebellar malformations occur as one component of a broader developmental brain

disorder.


68

P25

Methyl-CPG binding protein 2 isoform-specific regulatory network in human cerebellum

Pejhan S.1, Olson C.O. 1, Del Bigio M. 2, Siu V. 3, Ang L.C. 4, Rastegar M. 1

1 Regenerative Medicine Program, and Department of Biochemistry and Medical

Genetics, College of Medicine, Faculty of Health Sciences, University of Manitoba, MB,

Canada; 2 Department of Pathology, College of Medicine, Faculty of Health Sciences,

University of Manitoba, MB, Canada, 3 Department of Biochemistry, Schulich School of

Medicine and Dentistry, Western University, ON, Canada; 4 Department of Pathology,

Schulich School of Medicine and Dentistry, Western University, ON, Canada

Rett syndrome (RTT) is a severe neurodevelopmental disorder that affects mainly

females who are apparently normal at birth, but develop delay and regression in their

developmental milestones after 6 to 18 months. Mutation in the Methyl-CPG binding

protein 2 gene (MECP2) is the principal cause of RTT, and motor deficits such as loss of

motor coordination, ataxia, and gait apraxia are among the common phenotypes of this

syndrome. Impairments in modulatory neurotransmitter systems have been shown in

small size cerebellum of RTT animal models, which correspond well with the motor

phenotype observed in these animals. MeCP2 level in rat brain has been shown to be

regulated by Brain Derived Neurotrophic Factor (BDNF), and miR132, which is a

neuronal-specific microRNA. Our lab has shown region- specific distribution of MeCP2

isoforms in mice brain, which is in concordance with not fully redundant function of the

isoforms suggested by different studies. Considering the conservation of miR132 from

rodents to humans, and similarities of BDNF structure among mammals, our project

investigates the regulatory association between MeCP2 isoforms, BDNF, and miR132 in

human cerebellum. This study clarifies how region-specific impairments in MeCP2

regulatory systems may be involved in the pathogenesis of RTT. In order to study MeCP2

homeostasis regulation in human cerebellum, we examine post-mortem cerebellar tissue

of RTT patients with mutations in three different functional units of MeCP2, in comparison

with sex and age-matched controls. The fresh frozen samples are studied for transcription

level of MeCP2 isoforms, BDNF, and miR132. The protein levels are assessed in frozen

and fixed samples. The regulatory mechanisms will be studied in human neurons derived

from neural stem cells.

The outcome of our research provides further insight into MeCP2 isoforms regulatory

network in human cerebellum, and how its deregulation may correlate with the motor

deficits in RTT patients.


69

P26

Embryonic cerebellar graft development in the cerebellum of normal and cerebellar

mutant mice

Cendelin, J. 1, 2, Purkartova. Z.1, Kubik, J.1, Ulbricht, E.1, Kolinko, Y.3, 4

1 Department of Pathological Physiology, Faculty of Medicine in Pilsen, Charles

University, Pilsen, Czech Republic; 2 Laboratory of Neurodegenerative Disorders, Biomedical Center, Faculty of Medicine in

Pilsen, Charles University, Pilsen, Czech Republic; 3 Laboratory of Quantitative Histology, Biomedical Center, Faculty of Medicine in Pilsen,

Charles University, Pilsen, Czech Republic; 4 Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles

University, Pilsen, Czech Republic.

Neurotransplantation therapy is investigated as a potential tool to treat cerebellar

degenerations and their consequences. For this purpose, various cerebellar mutant mice

are used as models of these diseases. The aim of this work was to study the effect of two

different mouse cerebellar degenerations and of the background mouse strain on survival

and morphology of embryonic cerebellar grafts.

Mouse embryonic (E12) cerebellar cell suspension was injected bilaterally into the

cerebellum of adult Lurcher mice of the B6CBA and C3H strains, Purkinje cell

degeneration (pcd) mice of the B6.BR strain and wild type mice of the same strains. The

mice were sacrificed 2 months (to compare Lurcher and pcd mice) or 6 months (to

compare B6CBA and C3H Lurcher mice) after the surgery and their brains were examined

histologically for the presence, morphology and volume of the graft.

The graft survived in the vast majority of mice of all types. In both B6CBA and C3H

Lurcher mice only few graft-derived cells were found outside the main graft mass and the

fibers sprouting from the graft were rare. On the other hand, in wild type mice and in pcd

mutants colonization of host’s molecular layer adjacent to the graft site by graft derived

Purkinje cell-shaped cells and fiber sprouting have been observed.

This study was supported by the Charles University Research Fund (project number Q39)

and by the National Sustainability Program I (NPU I) Nr. LO1503 provided by the Ministry

of Education Youth and Sports of the Czech Republic.


70


Theme 5- Making Connections/Synaptogenesis in Cerebellum.

P27 / Prof. Naofumi Uesaka / Roles of retrograde signaling in climbing fiber to Purkinje cell

synapse elimination during postnatal cerebellar development.

P28 / Mr. Gideon Sarpong/ Compartmental organization in the mouse cerebellar nuclei and

inferior olive based on molecular expression patterns, and topography of Purkinje cell and

climbing fiber axonal projections.

P29 / Dr. Hirofumi Fujita/ Molecularly and anatomically distinct types of projection neurons in

the medial cerebellar nucleus mediate diverse outputs of the cerebellar vermis.

P30 / Dr. Aasef Shaikh/ Impaired motor learning in a disorder of the inferior olive: the

cerebellum learns a bad habit.


71

P27

Roles of retrograde signaling in climbing fiber to Purkinje cell synapse elimination during

postnatal cerebellar development

Uesaka N. 1, Abe M. 2, Yamazaki M. 2, Konno K. 3, Mikuni T. 1, Watanabe M. 3,

Sakimura K. 2 and Kano M. 1

Elimination of early-formed redundant synapses during postnatal development is

essential for functional neural circuit formation. Purkinje cells (PCs) in the neonatal

cerebellum are innervated by multiple climbing fibers (CFs). During postnatal

development, a single CF is selectively strengthened in each PC and becomes a ‘winner’

CF that is presumed to remain into adulthood, whereas the other ‘loser’ CFs are

eliminated. These developmental changes are dependent on postsynaptic activity in PCs.

Our recent studies have uncovered molecular mechanisms by which postsynaptic PCs

regulate CF synapse elimination. We have demonstrated that Semaphorin7A and

Semaphorin3A mediate retrograde signals from postsynaptic PCs to presynaptic CFs and

regulate elimination and maintenance respectively of CF synapses (Uesaka et al.,

Science, 2014). We further screened candidate retrograde signaling molecules for

strengthening or eliminating of CF synapses. We found that PC-specific knockout or

knockdown of progranulin, a growth factor implicated in the pathogenesis of

frontotemporal dementia, accelerated elimination of redundant CFs and reduced the

amplitude of synaptic inputs from winner CFs from postnatal day 11 (P11) to 16. These

results suggest that, similarly to Semaphorin3A, progranulin retrogradely

strengthens/maintains CF synapses and counteracts their elimination. Knockdown of

Semaphorin3A in progranulin-deficient PCs induced larger changes than those by

progranulin deletion alone, suggesting that progranulin and Semaphorin3A function

independently and may cooperate to maintain CF synapses. Furthermore, we found that

knockdown of Sort1, which is known to bind to progranulin, in CFs resulted in the similar

phenotypes to those caused by the progranulin deletion in PCs. These results suggest

that progranulin derived from PCs retrogradely acts on Sort1 in CFs,

strengthens/maintains CF synapses and thus counteracts CF synapse elimination from

P11 to P16.


72

P28

Compartmental organization in the mouse cerebellar nuclei and inferior olive based on

molecular expression patterns, and topography of Purkinje cell and climbing fiber axonal

projections

Sarpong G.A. 1, Fujita H. 1, 2, Vibulyaseck S. 1, Sugihara I. 1

1 Department of Systems Neurophysiology, Tokyo Medical and Dental University

Graduate School, Bunkyo-ku, Tokyo, Japan; 2 Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University

School of Medicine, Baltimore, Maryland, U.S.A.

The adult cerebellum is organized into longitudinal compartments that are revealed by

specific axonal projections and thus are presumably involved in different functions.

Heterogeneous Purkinje cell (PC) subsets with different molecular compositions are

distributed into transverse zones (combination of lobules) and parasagittal stripes in the

cerebellar cortex to form multiple compartments. In this study, we compared the

differences in expression pattern of some heterogeneously-expressed molecules,

including Protocadherin 10 (Pcdh10), with aldolase C (=zebrin II), which has been the

most extensively used marker for compartmentalizing the cerebellum, to identify

parasagittal stripes of PC subsets finer than those identified solely by aldolase C.

The Pcdh10 expression pattern was visualized by the reporter molecule β-galactosidase

using heterozygote samples of the knock-in mouse strain OL-KO, and analyzed in the

cerebellar cortex, nuclei (CN) and inferior olive (IO). In addition, we investigated PC and

climbing fiber axonal projections of cortical subpopulations using tracer injection.

Results of β-gal histochemistry revealed that Pcdh10 can selectively label PCs within the

cerebellar cortex by identifying distinct compartments and sub-stripes in other areas

particularly the paraflocculus where aldolase C does not show clear patterning. Pcdh10

was, in addition expressed in topographically matching subareas in the cerebellar nuclei

and IO. Each injection of tracer in the cortex labeled neurons or axonal terminals in a

single sub-nucleus of the IO or CN, indicating presence of fine compartmentalization. We

identified subareas that are topographically connected to different stripes in the

paraflocculus (distinguished by Pcdh10 expression), in the ventral part of the cerebellar

nucleus and ventrolateral part of the inferior olive. This heterogeneous expression of

Pcdh10 in PC subsets may be linked with topographic axonal projections and formation

of neuronal circuits.


73

P29

Molecularly and anatomically distinct types of projection neurons in the medial

cerebellar nucleus mediate diverse outputs of the cerebellar vermis

Fujita H. 1, Sima RJ. 1, Kodama T. 1, du Lac S. 1.

1Johns Hopkins University, Baltimore, USA.

The cerebellar vermis of the ‘spinocerebellum’ has been suggested to play critical roles

not only in motor control but also in limbic and cognitive functions. The neural substrates

for the diverse vermal functions, however, have not been fully understood due to the lack

of molecular markers for cell-type specific manipulation and the divergent projections from

its output nucleus, the medial cerebellar (fastigial) nucleus (FN). We hypothesized that

the fastigial neurons projecting to distinct downstream targets belong to molecularly

distinct neuronal types. To establish fastigial neuronal types, we performed single-cell

gene expression profiling with acutely dissociated fastigial neurons, with a focus on ion

channels and cell type marker candidates. The distribution of identified FN cell type was

subsequently examined by immunostaining for identified marker molecules. Projection

targets of each cell type were identified with anterograde and retrograde tracing combined

with immunostaining for marker molecules. Overall we discovered four major excitatory

cell types in the FN, that were distinguished by molecular expression, distribution within

the FN, and projection targets: 1) large Spp1+ neurons in the rostral FN and dorsolateral

protuberance projecting to vestibular nuclei, medullarly reticular formation, and spinal

cord, 2) small Calb2+ neurons in the ventrolateral FN projecting to vestibular nuclei, 3)

small to medium-sized Spp1+ and Snca+ neurons in the caudal FN projecting to VL

thalamus, superior colliculus, mesencephalic/pontine/medullary reticular formation,

vestibular nuclei, and spinal cord, and 4) small Snca+ neurons in the caudal FN projecting

to intralaminar and VM thalamus, hypothalamus, and brainstem neuromodulatory nuclei.

These results revealed that the cerebellar vermis broadcasts its output to a variety of

brain regions through at least four FN cell types, each of which may play a specific role

in the diverse functions of the cerebellar vermis.


74

P30

Impaired motor learning in a disorder of the inferior olive: the cerebellum learns a bad

habit

Aasef G. Shaikh 1, Aaron Wong 2, Lance M. Optican 3, David S. Zee 2

1 Case Western Reserve University, Cleveland, USA 2 Johns Hopkins University, Baltimore, USA 3 NIH, Bethesda, USA

Objective: An attractive hypothesis about how the brain learns to keep its motor

commands accurate is centered on the idea that the cerebellar cortex associates error

signals carried by climbing fibers with simultaneous activity in parallel fibers. Motor

learning can be impaired if the error signals are not transmitted, are incorrect, or are

misinterpreted by the cerebellar cortex. Learning might also be impaired if the brain is

overwhelmed with a sustained barrage of meaningless information unrelated to

simultaneously appearing error signals about incorrect performance. We demonstrate

that spontaneous, synchronous, and meaningless output from the inferior olive interferes

with the motor learning by disrupting or masking appropriate errors signals destined for

the cerebellum.

Methods: We test this concept in patients with syndrome of oculopalatal tremor (OPT), a

rare disease with spontaneous and irregular oscillations of the eyes. Such oscillations are

thought to reflect an abnormal, synchronous, spontaneous discharge to the cerebellum

from the degenerating neurons in the inferior olive.

Results and Conclusions: We examined motor learning during a short-term saccade

adaptation paradigm in patients with OPT and found a unique pattern of disturbed

adaptation, very different from the abnormal learning from direct involvement of the

cerebellum. In the OPT patients both fast (seconds) and slow (minutes) timescales of

learning were impaired. We suggest that the spontaneous, continuous, synchronous

output from the inferior olive prevents the cerebellum from receiving the error signals it

needs for appropriate motor learning. The important message from this study is that the

dysmetria due to impairment in motor learning is not exclusive feature of cerebellar

disorders, but it also highlights disorders of the inferior olive and its connections to the

cerebellum.


75


Theme 6- Aberrations of cerebellar Development and Function: Motor and Treatment

models.

P31 / Dr. Jens Claassen/ Cerebellar tDCS effects on the adaptation of arm reaching movements

to force-field and visuomotor perturbations.

P32 / Mrs. Marie Beaudin/ Systematic review of autosomal recessive ataxias and proposal for a

classification.

P33 / Dr. Lauren Watson/ Using induced pluripotent stem cells to investigate the disease

mechanisms of spinocerebellar ataxia.

P34 / Miss. Kim van Dun/ Targeting the cerebellum by noninvasive neurostimulation: a review.

P35 / Dr. Chiara Ferrari/ Visual motion discrimination and the vermis: A pilot study with slow-

frequency TMS.

P36 / Dr. Thais Monte/ Validation of the Neurological Examination Score for the assessment of

Spinocerebellar Ataxias (NESSCA) and responsiveness of several rating scales in

Spinocerebellar Ataxia type 2

P37 / Dr. Thais Monte/ Neurological phenotypes in spinocerebellar ataxia type 2: role of

mitochondrial polymorphism A10398G and other risk factors.

P38 / Prof. Laura Jardim/ The progression rate of neurological deterioration in spinocerebellar

ataxia type 2 changes according to stage of disease.

P39 / Prof. Laura Jardim/ Prediction of the age at onset in Spinocerebellar ataxia type 3 varies

according to population of origin.

P40 / Prof. Carlo Casali/ Gait abnormalities in Patients with Degenerative Cerebellar Ataxias.

P41 / Prof. Hong Jiang/ Alteration of methylation status in the ATXN3 gene promoter region is

linked to the SCA3/MJD.

P42 / Prof. Hong Jiang/ Association study between CAG repeats of PolyQ-related genes and

SCA3/MJD.

P43 / Prof. Hong Jiang/ Ubiquitin-related network underlain by (CAG)n loci modulate age at

onset in Machado-Joseph disease.

P44 / Prof. Hong Jiang/ ATXN2 polymorphism modulates age at onset in spinocerebellar ataxia

3/Machado-Joseph disease.

P45 /Dr. Basma Yacoubi/ SCA6 with stereotypical time control: evidence for impaired

functional capacity and differential neural activation.


76

P46 /Dr. David Szmulewicz/ A novel oculomotor biomarker in Friedreich’s Ataxia.

P47 /Dr. David Szmulewicz/ Toward objective clinical diagnosis of cerebellar ataxia.

P48 /Dr. David Szmulewicz/ Developing a clinically meaningful instrumented measure of upper

limb function in Friedreich ataxia.


77

P31

Cerebellar tDCS effects on the adaptation of arm reaching movements to force-field and

visuomotor perturbations

Claassen J 1, Mamlins A1, Hulst T1, Donchin O 2, Timmann D 1

1University Duisburg-Essen, Department of Neurology, Essen, Germany; 2Ben-Gurion University of the Negev, Department of Biomedical Engineering and

Zlotowski Center for Neuroscience, Beer Sheva, Israel.

Introduction: Anodal transcranial direct current stimulation (tDCS) of cerebellar

hemispheres leads to faster adaptation of arm reaching movements to visuomotor

rotation and force field perturbations (Galea et al., 2011; Herzfeld et al., 2014). The aim

of the present study was to replicate these findings and, secondly, to investigate the

influence of the onset of tDCS stimulation.

Patients and Methods: 120 healthy and right-handed subjects (60 females, aged 18-31

years, mean 23.2 ± 2.7) had to moved a cursor with a two-joined manipulandum to one

of eight targets presented on a screen. There were three baseline blocks, one adaptation

block and three washout blocks. 60 subjects performed a force field task (velocity-

dependent clockwise perpendicular force), 60 subjects a visuomotor adaptation task (30°

clockwise rotation). Equal numbers of subjects received anodal, cathodal or sham

stimulation over the right cerebellum beginning in the third baseline or the adaptation

block. The maximum error, final error and perpendicular velocity were assessed.

Results: During force field and visuomotor adaptation all subjects showed significant

effects of learning. They improved significantly regarding maximum error, final error and

perpendicular velocity (p < 0.001). There were no significant effects of stimulation, no

significant effects of onset of stimulation and no significant interactions. The same was

true for the washout blocks (all p-values > 0.05).

Conclusion: We were unable to replicate previous findings of modulatory cerebellar tDCS

effects. Furthermore, no differential effects of the onset of stimulation were observed. Our

results show that prior possible clinical application, future experiments are needed to

determine which cerebellar tDCS and task parameters lead to robust tDCS effects.

Funded by ELAN and DFG TI 239/16-1


78

P32

Systematic review of autosomal recessive ataxias and proposal for a classification

Beaudin M. 1, Klein C.J. 2, Rouleau G.A. 3, Dupré N. 1, 4

1 Faculty of Medicine, Université Laval, Quebec city, Canada 2 Department of Neurology, Mayo Clinic, Rochester, USA 3 Department of Neurology and Neurosurgery, McGill University, Montreal, Canada 4 Department of Neurological Sciences, CHU de Quebec - Université Laval, Quebec

city, Canada

BACKGROUND The classification of autosomal recessive ataxias represents a

significant challenge because of high genetic heterogeneity and complex phenotypes.

We conducted a comprehensive systematic review of the literature to examine all

recessive ataxias in order to propose a new classification and properly circumscribe this

field as new technologies are emerging for comprehensive targeted gene testing.

METHODS: We searched Pubmed and Embase to identify original articles on recessive

forms of ataxia in humans for which a causative gene had been identified. Reference lists

and public databases, including OMIM and GeneReviews, were also reviewed. We

evaluated the clinical descriptions to determine if ataxia was a core feature of the

phenotype and assessed the available evidence on the genotype-phenotype association.

Included disorders were classified as primary recessive ataxias, as other complex

movement or multisystem disorders with prominent ataxia, or as disorders that may

occasionally present with ataxia.

RESULTS After removal of duplicates, 2354 references were reviewed and assessed for

inclusion. A total of 130 articles were completely reviewed and included in this qualitative

analysis. The proposed new list of autosomal recessive ataxias includes 45 gene-defined

disorders for which ataxia is a core presenting feature. We propose a clinical algorithm

based on the associated symptoms.

CONCLUSION: We present a new classification for autosomal recessive ataxias that

brings awareness to their complex phenotypes while providing a unified categorization of

this group of disorders. This review should assist in the development of a consensus

nomenclature useful in both clinical and research applications.


79

P33

Using induced pluripotent stem cells to investigate the disease mechanisms of

spinocerebellar ataxia

Watson L.M. 1, Wong M.M.K. 1, Cowley S.A. 2, Vowles J. 2, Pribadi M. 3, Coppola G. 3,

Fogel B.L. 3, Becker E.B.E. 1

1 Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford,

United Kingdom 2 Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom 3 Department of Neurology, University of California at Los Angeles, Los Angeles,

California, USA

The spinocerebellar ataxias (SCAs) are a heterogeneous group of incurable brain

diseases, defined by a loss of motor coordination. The vast number of SCA-causing

genes identified to date presents a challenge to understanding disease mechanisms and

developing therapies. However, increasing evidence points towards common pathogenic

pathways shared across multiple SCA subtypes, which could provide novel therapeutic

targets. Through our work, Transient Receptor Potential Cation Channel, Subfamily C,

Member 3 (TRPC3), a non-selective cation channel, has emerged as a central player in

cerebellar ataxia. TRPC3 is required for metabotropic glutamate receptor subtype 1

(mGluR1)-dependent synaptic transmission in cerebellar Purkinje cells. Defects in

TRPC3 and mGluR1 signalling are linked to several genetically distinct forms of ataxia,

including SCA1, 3, 5 and 14, making TRPC3-mediated signalling a strong candidate for

a common ataxia pathway. We have also recently identified the first human ataxia patient

harbouring a mutation (R762H) in the TRPC3 gene, causing a novel disorder now termed

SCA41 (OMIM #616410). Overexpression of mutant TRPC3 in Neuro-2A cells resulted in

increased cell death and enhanced calcium signalling, consistent with increased channel

activity, similar to the mouse Mwk Trpc3 mutation that causes ataxia via a toxic gain-of-

function mechanism. To investigate the role of TRPC3 in a more physiologically relevant

model of human ataxia, we have developed a novel method for the differentiation of

cerebellar neurons from induced pluripotent stem cells (iPSCs). Using this approach, we

have generated the first SCA41 patient Purkinje cells in vitro, which show characteristic

dendritic branching and stain positive for Calbindin-D28k. Current investigations are

focused on the evaluation of gene expression patterns, survival, morphology and

electrophysiology in these cells, in order to identify pathogenic mechanisms which may

be relevant to multiple subtypes of SCA.


80

P34

Targeting the cerebellum by noninvasive neurostimulation: a review

Van Dun K 1, Mariën P 1, 2, Manto M 3, 4

1 Clinical and Experimental Neurolinguistics, Vrije Universiteit Brussel, Brussels,

Belgium 2 Department of Neurology and Memory Clinic, ZNA Middelheim General Hospital,

Antwerp, Belgium 3 Unité d’Etude du Mouvement, Laboratoire de Neurologie Expérimentale, Université

libre de Bruxelles (ULB), Brussels, Belgium 4 Service des Neurosciences, Université de Mons, Mons, Belgium

Improving brain functions by modulating neuronal excitability with noninvasive techniques

such as tDCS and TMS is an exciting new research domain. Since the cerebellum is

intrinsically connected with the cortical regions subserving motor, cognitive and affective

functions, it might be an interesting target. We conducted a literature search to determine

the validity of cerebellar stimulation in each of these domains.

The cerebellum is involved in motor functioning and is responsible for monitoring ongoing

movements, predicting future states, and detecting and correcting errors (state

estimations). Cerebellar TMS and tDCS seem to have a different impact on motor

functioning depending on complexity, task, and strategy. Cerebellar stimulation usually

interferes with cognitive processing in a subtle but definite manner. Therefore, specific

methods to measure potential effects and timing are crucial. A lot of parallels may be

drawn with the motor literature, with a differential impact depending on complexity, task,

and strategy, and a role for the cerebellum in perception/processing, error correction,

learning, and accuracy. Only a handful of studies have investigated the impact of

cerebellar stimulation on affective processing but there is evidence that cerebellar

stimulation might modulate affective and somatosensory processing.

In clinical populations, cerebellar TMS and tDCS might become powerful substituting or

adjuvant therapeutic tools. Restoration of the cerebello-cerebral functional connectivity

by means of cerebellar stimulation might not only improve motor but also nonmotor

cognitive and affective deficits following cerebellar pathology. Several studies have

shown that repeated sessions of cerebellar stimulation may exert long-lasting positive

effects. However, in order to use TMS and tDCS as standard (clinical) practice

techniques, it is crucial to learn more about the working mechanisms and impact of the

different stimulation protocols.


81

P35

Visual motion discrimination and the vermis: A pilot study with slow-frequency TMS

Ferrari C. 1, Vecchi T. 2, 3, Oldrati V. 2, 3, Reverberi C. 1, & Cattaneo Z. 1, 3

1 Department of Psychology, University of Milano-Bicocca, Milan 20126, Italy. 2 Department of Brain and Behavioral Sciences, University of Pavia, Pavia 27100, Italy. 3 Brain Connectivity Center, National Neurological Institute C. Mondino, Pavia 27100,

Italy.

A new hypothesis suggesting a possible role of the cerebellum in coordinating sensory

data in the motor domain and in the cognitive domain has been proposed. However,

inconsistent evidence exists relatively to the topographic contribution of the cerebellum

to this function, with some lesion and neuroimaging studies pointing to the midline

structures but not to cerebellar hemispheres and others pointing also to the lateral parts

of the cerebellum as crucial in underpinning motion discrimination. A recent study showed

participants' impaired performance in identifying the direction of moving dots while

transcranial magnetic stimulation (TMS) was delivered over the vermis but not while TMS

was delivered over the hemispheres. The aim of the present study is to extend prior

evidence relative to the contribution of the vermis to motion perception processes, by

employing more complex stimuli, including dots moving on the horizontal axis (leftward

and rightward) and dots moving on the vertical axis (downward and upward). We asked

participants to perform a motion discrimination task before and after receiving TMS over

the vermis, V1 and a control condition (vertex). We found that TMS applied over the

vermis significantly impaired participants’ performance, similarly to TMS over V1,

corroborating previous findings on the causal role of the vermis in motion perception.

Moreover, the detrimental effects of TMS over the cerebellum were comparable when

dots moved on the horizontal axis and on the vertical axis. Results are discussed in light

of sensory integration mechanisms possibly contributing to this phenomenon. Finally, our

findings support the possible employment of TMS as a tool to promote changes in the

brain activity paralleled by behavioral modifications able to overcome the exact time of

the stimulation.


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P36

Validation of the Neurological Examination Score for the assessment of Spinocerebellar

Ataxias (NESSCA) and responsiveness of several rating scales in Spinocerebellar

Ataxia type 2

Monte TL 1,7 , Reckziegel ER 9, Augustin MC 9, Silva ASP 9, Locks-Coelho LD 9,

Barsottini OP 10, Pedroso JL 10, Vargas FR 11,12, Saraiva-Pereira ML 2, 3, 6, Leotti VB 4,8,

Jardim LB 2, 3, 5, 7, 8, 13, on behalf of Rede Neurogenética.

1 Serviço de Neurologia, Hospital de Clínicas de Porto Alegre, Rio Grande do Sul,

Brazil; 2 Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Rio Grande

do Sul, Brazil; 3 Laboratório de Identificação Genética, Hospital de Clínicas de Porto

Alegre, Rio Grande do Sul, Brazil; 4 Departamento de Matemática e Estatística,

Universidade Federal do Rio Grande do Sul, Brazil; 5 Departamento de Medicina

Interna, Universidade Federal do Rio Grande do Sul, Brazil; 6 Departamento de

Bioquímica, Universidade Federal do Rio Grande do Sul, Brazil;7 Programa de Pós-

Graduação em Ciências Médicas, Universidade Federal do Rio Grande do Sul, Brazil; 8

Programa de Pós-Graduação em Epidemiologia, Universidade Federal do Rio Grande

do Sul, Brazil; 9 Faculdade de Medicina, Universidade Federal do Rio Grande do Sul,

Brasil; 10 Setor de Neurologia Geral e Ataxias. Disciplina de Neurologia Clínica da

UNIFESP - Escola Paulista de Medicina, Universidade Federal de São Paulo, Brazil;11

Laboratório de Epidemiologia de Malformações Congênitas, Fundação Oswaldo Cruz,

Rio de Janeiro, Brazil;12 Departamento de Genética e Biologia Molecular, Universidade

Federal do Estado do Rio de Janeiro, Brazil;13 Instituto Nacional de Genética Médica

Populacional, Brazil.

Background: Spinocerebellar ataxia type 2 (SCA2), caused by a CAG expansion

(CAGexp) at ATXN2, has a complex clinical picture. While validated ataxia scales are

available, comprehensive instruments to measure all SCA2 neurological manifestations

are required. Aim: to validate the Neurological Examination Score for the assessment of

Spinocerebellar Ataxias (NESSCA) to be used in SCA2 and to compare its

responsiveness to those obtained with other instruments.

Methods: NESSCA, SARA, SCAFI, and CCFS scales were applied in symptomatic SCA2

patients. Correlations were done with age at onset, disease duration, CAGexp, and

between scales. Responsiveness was estimated by comparing deltas of stable to worse

patients after 12 months, according to Patient Global Impression of change, and the area

under the curve (AUC) of the Receiver Operating Characteristics curve of scores range.

Results: Eighty-eight evaluations (49 patients) were obtained. NESSCA had an even

distribution, and correlated with disease duration (r=0.55), SARA (r=0.63), and CAGexp

(rho=0.32): both explained 44% of NESSCA variance. Deltas (CI 95%) after one year in


83

stable and worse patients were only significantly different for SARA. NESSCA, SARA,

SCAFI, and CCFS AUC were 0.63, 0.81, 0.49, and 0.48, respectively.

Discussion: NESSCA is valid to be used in SCA2. However, the only instrument that

presented good responsiveness to change in one year was SARA. We suggest that

NESSCA can be used as a secondary outcome in future trials in SCA2, due to the burden

of neurological disabilities related to disease progression.

P37

Neurological phenotypes in spinocerebellar ataxia type 2: role of mitochondrial

polymorphism A10398G and other risk factors

Monte TL 1,6 , Pereira FS 8, Reckziegel ER 3, Augustin MC 3, Silva ASP 3, Locks-Coelho

LD 3, Pedroso JL 10, Barsottini OP 10, Vargas FR 11, Saraiva-Pereira ML 2,4,7,8,9, Jardim

LB 1, 2, 3, 5, 7, 8, 9,12, on behalf of Rede Neurogenética.

1 Programa de Pós-Graduação em Ciências Médicas, Universidade Federal do Rio

Grande do Sul, Brazil; 2 Programa de Pós-Graduação em Genética e Biologia

Molecular, Universidade Federal do Rio Grande do Sul, Brazil; 3 Faculdade de

Medicina, Universidade Federal do Rio Grande do Sul, Brazil; 4 Departamento de

Bioquímica, Universidade Federal do Rio Grande do Sul, Brazil; 5 Departamento de

Medicina Interna, Universidade Federal do Rio Grande do Sul, Brazil; 6 Serviço de

Neurologia, Hospital de Clínicas de Porto Alegre, Rio Grande do Sul, Brazil; 7 Serviço

de Genética Médica, Hospital de Clínicas de Porto Alegre, Rio Grande do Sul, Brazil;8

Laboratório de Identificação Genética, Hospital de Clínicas de Porto Alegre, Rio Grande

do Sul, Brazil;9 Rede Neurogenética, Hospital de Clínicas de Porto Alegre, Rio Grande

do Sul, Brazil; 10 Setor de Neurologia Geral e Ataxias. Disciplina de Neurologia Clínica

da UNIFESP - Escola Paulista de Medicina, Universidade Federal de São Paulo,

Brazil;11 Departamento de Genética e Biologia Molecular, Universidade Federal do

Estado do Rio de Janeiro, Brazil;12 Instituto Nacional de Genética Médica Populacional,

Brazil.

Background: Spinocerebelar ataxia type 2 (SCA2) is due to a CAG expansion (CAGexp)

at ATXN2. Alongside characteristic ataxia with saccadic slowness, SCA2 presents great

clinical variability. Aims: to study parkinsonism, dementia, dystonia, and amyotrophy, as

subphenotypes of SCA2, and to explore the effect of CAG repeats at several loci and of

mitochondrial polymorphism A10398G as modifiers of phenotype.

Methods: Symptomatic subjects were classified by presence/absence of neurological

signs mentioned above; SARA and NESSCA scores were obtained. CAG repeats at


84

ATXN1, ATXN2, ATXN3, CACNA1A, ATXN7 and RAI1, and polymorphism A10398G at

mtDNA were established. Group characteristics were compared, with a p < 0.05.

Results: Forty-eight SCA2 individuals were included. Age at onset, CAGexp, and disease

duration explained 53% and 43% of SARA and NESSCA variations. CAGexp of subjects

with and without parkinsonism were different (medians of 42 and 39 repeats) as well as

of subjects with and without dystonia (44 and 40 repeats). Amyotrophy was not

significantly related to any variable under study. Concerning polymorphism A10398G,

83% of subjects with and 34% of those without cognitive decline carried 10398G at

(p=0.003).

Discussion: Treating the four phenotypical subgroups as oucomes was a valid strategy

to identify modifiers of disease. Among the correlations found, some confirmed previous

reports, such as those between dystonia and CAGexp. Of note was the association

between cognitive decline and the variant G at mitochondrial polymorphism A10398G, a

variant formerly related to earlier ages at onset in SCA2.

P38

The progression rate of neurological deterioration in spinocerebellar ataxia type 2

changes according to stage of disease

Monte TL1, 7 , Reckziegel ER 10, Augustin MC 10, Locks-Coelho LD 10, Silva ASP 10,

Barsottini OP 11, Pedroso JL 11, Vargas FR 12, 13, Saraiva-Pereira ML 2, 6, 8 , Camey SA 4

,9, Leotti VB 4, 9, Jardim LB 2, 3, 5, 7, 8, 10, 14, on behalf of Rede Neurogenética.

1 Serviço de Neurologia, Hospital de Clínicas de Porto Alegre, Rio Grande do Sul,

Brazil; 2 Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Rio Grande

do Sul, Brazil; 3 Laboratório de Identificação Genética, Hospital de Clínicas de Porto

Alegre, Rio Grande do Sul, Brazil; 4 Departamento de Estatística, Universidade Federal

do Rio Grande do Sul, Brazil; 5 Departamento de Medicina Interna, Universidade

Federal do Rio Grande do Sul, Brazil; 6 Departamento de Bioquímica, Universidade

Federal do Rio Grande do Sul, Brazil;7 Programa de Pós-Graduação em Ciências

Médicas, Universidade Federal do Rio Grande do Sul, Brazil; 8 Programa de Pós-

Graduação em Genética e Biologia Molecular , Universidade Federal do Rio Grande do

Sul, Brazil; 9 Programa de Pós-Graduação em Epidemiologia, Universidade Federal do

Rio Grande do Sul, Brazil; 10 Faculdade de Medicina, Universidade Federal do Rio

Grande do Sul, Brasil; 11 Setor de Neurologia Geral e Ataxias. Disciplina de Neurologia

Clínica da UNIFESP - Escola Paulista de Medicina, Universidade Federal de São

Paulo, Brazil;12 Laboratório de Epidemiologia de Malformações Congênitas, Fundação

Oswaldo Cruz, Rio de Janeiro, Brazil;13 Departamento de Genética e Biologia


85

Molecular, Universidade Federal do Estado do Rio de Janeiro, Brazil; 14 Instituto

Nacional de Genética Médica Populacional, Brazil.

Background: Spinocerebellar ataxia type 2 (SCA2) has heterogeneous symptoms.

Previous studies showed progression of ataxic manifestations only, and all used the study

entry as the start of the measurements. Aims: to describe the progression of Scale for the

Assessment and Rating of Ataxia (SARA), SCA Functional-Index (SCAFI), Composite-

Cerebellar-Functional-Score (CCFS), and the Neurological Examination Score for

Spinocerebellar Ataxias (NESSCA) in SCA2; to explore whether progression is linear

during all the disease duration; and to look for potential modifiers.

Methods: 49 subjects were examined. Age at onset and disease duration, CAGexp, and

amyotrophy, parkinsonism, dystonia, and cognitive losses at baseline were used as

independent variables. Linear growth curve models were adjusted to model relationships

between outcomes and time in two ways: a study duration model (baseline and follow up

observations) versus a disease duration model (disease onset according to patient,

baseline, and follow up observations).

Results: SARA progressed 1.75 versus 0.79 points/year in the study duration and disease

duration models. NESSCA progressed 1.45 versus 0.41 points/year in the study duration

and disease duration models. Therefore, NESSCA and SARA progression rates were not

constant during disease duration. Individuals with less and more than 10 years of disease

duration progressed 0.35 and 2.45 points/year in SARA scores (p = 0.013) in the study

duration model.

Discussion: Early phases of disease were associated with slower SARA and NESSCA

progressions. Modelling of future studies should take those parameters into account. Our

database was made available online in order to help future meta-analyses intended to

clarify SCA2 progression.

P39

Prediction of the age at onset in spinocerebellar ataxia type 3 varies according to

population of origin

Leotti VB 1,8, Mattos EP 2,7, Souza GN 3, Furtado GV 2,7, Saraiva-Pereira ML 2,4,6,7 ,

Saute JAM 6, Camey SA 1,8, Jardim LB 2,3,5,6

1 Departamento de Estatística, Universidade Federal do Rio Grande do Sul, Brazil.2

Programa de Pós-Graduação em Genética e Biologia Molecular, Universidade Federal

do Rio Grande do Sul, Brazil.3 Programa de Pós-Graduação em Ciências Médicas,

Universidade Federal do Rio Grande do Sul, Brazil.4 Departamento de Bioquímica,

Universidade Federal do Rio Grande do Sul, Brazil.5 Departamento de Medicina


86

Interna, Universidade Federal do Rio Grande do Sul, Brazil.6 Serviço de Genética

Médica, Hospital de Clínicas de Porto Alegre, Brazil.7 Laboratório de Identificação

Genética, Hospital de Clínicas de Porto Alegre, Brazil.8 Programa de Pós-Graduação

em Epidemiologia, Universidade Federal do Rio Grande do Sul, Brazil.

Background: The CAG repeat expansion (CAGexp) correlates with age at onset (AO) in

spinocebellar ataxia type 3/Machado-Joseph disease (SCA3/MJD). Recently, a specific

formula was proposed to predict AO based on CAGexp, based on an European

population (Tezenas du Montcel et al, 2014). Aim: to test whether the formula proposed

by the European study can predict accurately AO of Brazilian SCA3/MJD patients and to

build a new one, addressed to this population.

Methods: data of all SCA3/MJD carriers, living in our region, were registered and kept

confidential. A survey was performed among affected individuals to confirm information

on AO of gait ataxia (AOga). The predicted median AOga from birth were calculated for

symptomatic individuals from the survey and for anonymous data about asymptomatic

individuals, based on his/her CAGexp value with a critical range (CR) from 5th to 95th

percentiles. Equations and estimates presented by Tezenas du Montcel et al (2014) were

implemented.

Results: 100 symptomatic and 47 asymptomatic SCA3/MJD carriers were studied.

Predicted AOga underestimated the actual AOga by 10.41 years (CI 9.01 - 11.80).

Underestimations were present in all individuals who received a predicted AOga of 25

years or more. Thirty one (66%) of the 47 asymptomatic carriers were still asymptomatic

in an age older than the predicted AOga, based on their CAGexp. A formula addressed

to our population was developed.

Discussion: The data obtained from the European cohort was not sufficient to propose a

model to predict age at onset for SCA3/MJD in general. These differences pointed to a

populational stratification effect operating in SCA3/MJD. Prediction of the AOga should

be modelled for each specific population of origin.


87

P40

Gait abnormalities in Patients with Degenerative Cerebellar Ataxias

Carlo Casali, Giorgia Chini, Carmela Conte, Martina Rinaldi, Alberto Ranavolo,

Christian Marcotull, Luca Leonardi, Gaia Fragiotta, Fabiano Bini, Gianluca Coppola,

Francesco Pierelli, Mariano Serrao

Objectives: In the present study, the progression of gait impairment in a group of patients

with primary degenerative cerebellar ataxias was observed over a period of 4 years.

Materials and Methods: A total of 30 patients underwent an initial gait analysis study, and

thereafter only 12 were evaluated because they completed the 2- and 4-year follow-up

evaluations. Time-distance parameters, trunk and joint range of motion (RoM), and

variability parameters (e.g., coefficients of variation) were measured at the baseline and

at each follow-up evaluation. The scale for the assessment and rating of ataxia (SARA)

was used to evaluate disease severity.

Results and conclusions: We found a significant increase in the SARA score at both the

2- and 4-year follow-up evaluations. Almost all the gait variables changed significantly

only at the 4-year follow-up. Particularly, we found a significant decrease in the step length

and in the hip, knee, and ankle joint RoM values and noted a significant increase in the

trunk rotation RoM and stride-to-stride and step length variability. Furthermore, a

significant difference in ankle joint RoM was found between spinocerebellar ataxia and

sporadic adult-onset ataxia patients, with the value being lower in the former group of

patients. Our findings suggest that patients with degenerative cerebellar ataxias exhibit

gait decline after 4 years from the baseline. Moreover, patients try to maintain an effective

gait by adopting different compensatory mechanisms during the course of the disease in

spite of disease progression.


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P41

Alteration of methylation status in the ATXN3 gene promoter region is linked to the

SCA3/MJD

Wang C. 1, Peng H. 1, Li J. 2, Jiang H. 1,2,3

1 Department of Neurology,Xiangya Hospital, Central South University, Changsha,

Hunan, 410008, P. R. China; 2 State Key Laboratory of Medical Genetics, Central South

University, Changsha, Hunan, 410078, P. R. China; 3 Key Laboratory of Hunan Province

in Neurodegenerative Disorders, Central South University, Changsha, Hunan, 410008,

P. R. China

Background: DNA methylation can regulate the repeats instability of disease-causing

repeats and modify the neurodegenerative diseases progression. In this study, blood

samples collected from SCA3/MJD patients vs. control were used to explore the potential

link of DNA methylation levels at ATXN3 gene promoter to the pathogenesis of

SCA3/MJD.

Methods: 141 SCA3/MJD patients from 71 families and 112 health controls were enrolled

in our study. Density gradient centrifugation was used to isolate PBMCs of all subjects.

The DNA methylation levels of ATXN3 promoter were detected by BSP. Recombinant

DNA technology with T-vector cloning followed by direct DNA sequencing was applied to

evaluate the CAG repeats size of ATXN3. To further understand the regulation of ATXN3

promoter activity by DNA methylation, cell based luciferase assays were carried out.

Results: Hypermethylation status was observed in the first CpG island of ATXN3

promoter. Predominantly non-methylated status was observed in the second CpG island

of ATXN3 promoter. Statistically significant difference was found when comparing the first

CpG island methylation levels between the SCA3/MJD patients and controls (82.61±3.97

vs 78.97±2.22, p=0.00). Higher DNA methylation levels were observed in the patients

with earlier age at onset than the patients with later age at onset who shared the same

CAG repeats size in the same family. Higher levels of DNA methylation were observed in

the parents from SCA3/MJD kindreds with an intergenerational CAG repeats instability

than those without intergenerational CAG repeats instability (86.04±2.87 vs 80.00±2.81,

p=0.00). In addition, the first CpG island of the ATXN3 promoter served as the main

regulation region of DNA methylation.

Conclusions: This is the first study of DNA methylation levels of ATXN3 promoter using

BSP technology. We found potential links between epigenetics and the CAG repeats

stability at ATXN3 as well as AAO of SCA3/MJD patients. These findings suggested that

an epigenetic change may contribute to the pathogenesis of the SCA3/MJD and provide

potential therapeutic targets for CAG repeats based diseases.


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P42

Association study between CAG repeats of PolyQ-related genes and SCA3/MJD

Chen Z. 1, Zheng C. 1, Long Z1, Tang B. 1, 2, 3 Jiang H.1, 2, 3

1 Department of Neurology, Xiangya Hospital, Central South University, Changsha,

Hunan, P.R. China;2 Key Laboratory of Hunan Province in Neurodegenerative

Disorders, Central South University, Changsha, Hunan, P.R. China;3 State Key

Laboratory of Medical Genetics, Central South University, Changsha, Hunan, P.R.

China.

Objective: To investigate other factors involved in the variability of age at onset (AO) for

Chinese Han SCA3/MJD patients.

Background: Machado-Joseph disease (MJD), also known as spinocerebellar ataxia type

3 (SCA3), is the most prevalent subtype of SCAs in China. Studies showed that AO is

negatively correlated with the number of CAG repeats, but only 50-70% of the variation

in AO is explained by CAG repeat length in ATXN3. We speculate that some CAG repeats

of PolyQ-related genes may modulate the AO for SCA3/MJD.

Methods: A total of 802 SCA3/MJD patients from mainland China were recruited. All

subjects had genetically determined CAG repeat expansion in ATXN3 by PCR

amplification and capillary electrophoresis. The participating subjects were genotyped for

9 other polymorphic (CAG)n-containing genes (including ATXN1, ATXN2, CACNA1A,

ATXN7, TBP, ATN1, IT15, KCNN3, RAI1). Analysis of the CAG repeats related to the AO

of SCA3/MJD were performed using ANCOVA and multiple regressive.

Results: The AO did not differ according to the gender in the subjects or their transmitting

parents. The longer allele of ATXN3 contributed to 56.9% variation of AO for SCA3/MJD.

The shorter allele of ATXN3 with CAG repeats <19 or ranging from 26 to 40, contributed

to 1.3% and 1.9% variation of AO respectively.The longer allele of ATXN3 interacting with

the shorter allele of ATXN1, the difference of IT15 alleles and the difference of KCNN3

alleles contributed to 0.4%, 0.7% and 0.8% variation of AO respectively. Subjects with an

intermediate ATXN2 allele ranging from 27 to 32, which contributed to 23.2% variation of

AO, had an earlier AO (about 2.48±1.58 years).The intermediate CACNA1A homozygous

alleles ranging from 9 to 17 in SCA3/MJD patients contributed to 4.8% variation of AO.

The shorter allele of ATXN7 with CAG repeats <10 or the longer allele ranging from 10 to

15 in SCA3/MJD patients contributed to 2.5% and 1.6% variation of AO respectively. The

longer allele of ATXN7 with CAG repeatsranging from 12 to 17 and the shorter allele (>10)

in SCA3/MJD individuals contributed to 3.8% variation. The shorter RAI1 allele with CAG

repeats12 or the longer allele ranging from 13 to 14contributed to 0.7% and 5.2% variation

of AO respectively.The longer allele of RAI1 with CAG repeatsranging from 13 to 14 and

the shorter allele (>11) in SCA3/MJD individuals contributed to 7.3% variation.


90

Conclusion: In Chinese Han population the CAG repeats in the longer allele of ATXN3

contribute to 56.9% variation of AO in SCA3/MJD. ATXN1, ATXN2, KCNN3, CACNA1A,

ATXN7 and RAI1 gene may modulate the AO of SCA3/MJD.

P43

Ubiquitin-related network underlain by (CAG)n loci modulate age at onset in Machado-

Joseph disease

Chen Z. 1, Wang C. 1, Tang b.1,2,3, Jiang H. 1,2,3


Hunan, P.R. China;2 Key Laboratory of Hunan Province in Neurodegenerative

Disorders, Central South University, Changsha, Hunan, P.R. China;3 State Key


China.

Objective: To investigate networkbased genetic factors involved in the variability of age

at onset (AO) for Chinese Han SCA3/MJD patients.

Background: Machado-Joseph disease (MJD), also known as spinocerebellar ataxia type

3 (SCA3), is the most prevalent subtype of SCAs in China. Studies showed that AO is

negatively correlated with the number of CAG repeats, but only 46-76%of the variation in

AO is explained by CAG repeat length in ATXN3. We speculate that on a network of

(CAG)n loci may modulate the AO for SCA3/MJD.

Methods: A total of 825 SCA3/MJD patients from mainland China were recruited. All

subjects had genetically determined CAG repeat expansion in ATXN3 by PCR

amplification and capillary electrophoresis. The participating subjects were genotyped for

7 other polymorphic (CAG)n-containing genes (including ATXN1, ATXN2, CACNA1A,

ATXN7, TBP, ATN1, HTT). Analysis of ubiquitin-related network related to the AO of

SCA3/MJD were performed using interacting network and statistical analysis.

Results: Analysis of polyQ-relatedinteracting network between ATXN3 and other polyQ-

disease geneswas performed to generate fourubiquitin C (UBC)-related genetic modules:

I) ATXN3,ATXN1andATXN2;II) ATXN3, ATXN1, ATXN2, HTT; III) ATXN3, ATXN1,

ATXN2 and ATXN7; and IV)ATXN3, ATXN1, ATXN2, HTT, ATXN7, CACNA1A, TBP and

ATN1.Theoverall effect of the fourmodules on AO wasobserved for module I (p=0.005)

and module II (p=0.01), suggesting a more robust association among ATXN3, ATXN1,

ATXN2 andHTT,than with ATXN6, ATXN7, TBP and ATN1. A “module impact score” was

constructedby summing the differences between the actual and average size of

(CAG)nfor each allele at each locus, and weighted that with their effect in our


91

cohort,suggesting a positive correlation of AO with each quartile of this module impact

score andaccounting for 1.8% of the variance of residual AO.

Conclusion: We propose genetic networks contributing to variation of AO in MJD in an

ubiquitin-dependent pathway, offering a new perspective for candidate gene analysis.

P44

ATXN2 polymorphism modulates age at onset in spinocerebellar ataxia 3/Machado-

Joseph disease

Ding D. 1, Li K. 1, Wang C. 1, Tang B. 1, 2, 3, Jiang H. 1,2,3,4


Hunan, P.R. China; 2 Key Laboratory of Hunan Province in Neurodegenerative

Disorders, Central South University, Changsha, Hunan, P.R. China; 3 State Key


China; 4 Xinjiang Medical University, Xinjiang, 830011, P.R. China.

Objective: Spinocerebellar ataxia type 3/Machado-Joseph disease is caused by the

abnormal expansion of CAG repeats within exon 10 of the ATXN3. The length of

expanded CAG repeats in ATXN3 is negatively related with age at onset (AAO), but it

could only explain 50-70% of its variability, emphasizing other potential factors may

contribute to the variability. ATXN2 has been implicated in neurodegeneration and the

length of longer CAG alleles in ATXN2 was recently proved associating with age at onset.

In this study, we tried to explore a functional polymorphism (rs7969300) in ATXN2 in

Chinese population to determine whether and how much it will explain the variability of

AAO in SCA3/MJD.

Methods: The CAG repeats in ATXN3 and ATXN2 were determined by capillary

electrophoresis and DNA sequencing with T-vector cloning.Genotypes of the SNP were

examined by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry

via MassARRAY system. Linear regression analysis was performed to explore the

association between rs12957023 and AAO of SCA3/MJD patients.

Results: A significant association between rs7969300 and AAO was observed.The AAO

of patients who carrying the rare GG genotype was almost 3 years earlier than AG and

AA genotypes after adjusting for the size of abnormal expanded CAG repeats in ATXN3

and excluding the effect of CAG repeats in ATXN2.

Conclusion: This study firstly demonstrated that age at onset could be moderated by

rs7969300, which may also indicate that other than ever-known CAG alleles, SNPs could

also be taken into consideration as a new potential component of genetic factors when

exploring the variability of age at onset in the future.


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P45

SCA6 with stereotypical time control: evidence for impaired functional capacity and

differential neural activation

Yacoubi B.1, Casamento-Moran A.1, Subramony S.H.2, Vaillancourt D.E.1, 2, and

Christou E.A.1

1 Department of Applied Physiology and Kinesiology, University of Florida 2 Department of Neurology, University of Florida

Spinocerebellar ataxia type 6 (SCA6) is a genetic disease causing cerebellar

degeneration. Our purpose was to determine the effect of motor output variability on the

functional capacity of SCA6 individuals. Seventeen individuals diagnosed with SCA6

(60.7 ± 9.6 yrs., 10 F) and 12 healthy controls (60.7 ± 9.6 yrs., 10 F) performed 50 goal

directed contractions with ankle dorsiflexion. The target force was 15% maximum and the

target time was 180 ms. We provided visual feedback of the force trajectory relative to

the target 3 s after each trial. We recorded EMG activity of the agonist muscle (Tibialis

Anterior; TA). We quantified the following: 1) SCA6 functional capacity using clinical

assessments scales; the International Cooperative Ataxia Rating Scale (ICARS) and the

Scale for the assessment and Rating of Ataxia (SARA); 2) Force and time dysmetria; 3)

Force and time endpoint variability; 4) TA EMG burst; 5) TA EMG burst variability. We

identified two groups of SCA6 patients distinguished by the degree of variability in time

endpoint (stereotypy). The first group (n=8) exhibited low variability in time (12-19%;

stereotypical). The second group (n=9) exhibited a degree of variability comparable to

healthy controls (21-40%; normal variability). The stereotypical group exhibited impaired

functional capacity relative to the normal variability group as assessed by ICARS (37.4 ±

5.45 vs. 22.9 ± 5.02; P<0.05) and SARA (14.7 ± 2.17 vs 9.1 ± 1.78; P<0.05). It also

exhibited greater force dysmetria (81.6 ± 17 vs. 42.3 ± 8.7; P<0.05). The stereotypy in

time control was associated with higher fMRI activity in cerebellar lobules IV and V

(R2=0.26; P<0.05) and more variable TA EMG burst duration (R2=0.26; P<0.05). Our

findings provide novel evidence that SCA6 with stereotypical time control exhibit impaired

functional capacity and force control as well as differential activation of the cerebellum

and muscle.


93

P46

A novel oculomotor biomarker in Friedreich’s Ataxia

Szmulewicz DJ. 1, 2, 3, MacDougall HG. 4, Storey GM. 5, Halmagyi GM. 6, Cremer P. 7,

Corben L. 8,9,10, Delatycki M. 8, 9, 10, 11

1 Balance Disorders and Ataxia Service, Royal Victorian Eye and Ear Hospital,

Melbourne, Australia, 2 Cerebellar Ataxia Clinic, Neuroscience Department,

AlfredHealth, Melbourne, Australia, 3 Florey Institute of Neuroscience and Mental

Health, Melbourne, Australia, 4 Faculty of Science, University of Sydney, Sydney,

Australia, 5 Medicine, Nursing and Health Sciences Department, Monash University, 6

Department of Neurology, Royal Prince Alfred Hospital, New South Wales, Australia, 7

Department of Neurology, Royal North Shore Hospital, New South Wales, Australia, 8

Bruce Lefroy Centre for Genetic Health Research, Murdoch Childrens Research

Institute, Parkville, Vic., Australia, 9 School of Psychological Sciences, Monash

University, Clayton, Australia, 10 Department of Paediatrics, University of Melbourne,

Parkville, Australia, 11 Victorian Clinical Genetics Service, Parkville, Vic., Australia.

Objective: To investigate the vestibulo-cerebellar interaction in Friedreich's ataxia (FA) to

further elucidate the neuro-otological manifestations of this disease and elucidate a

possible bio-marker for FA clinical treatment trials.

Background: Friedreich's ataxia (FA) is the most commonly occurring inherited ataxia,

and involves widespread neurodegenerative sequelae. Whilst oculomotor, vestibular and

cerebellar affects have been documented, little is understood about the clinical

consequences of pathology affecting these interacting systems. Impairment of the visually

enhanced vestibulo-ocular reflex (VVOR; also called the “doll's head”, “doll's eye” or

oculo-cephalic reflex) reveals a compound deficit in the three compensatory reflexes

involved in eye movement, namely the vestibulo-ocular reflex, smooth pursuit, and the

optokinetic reflex.

Materials and methods: A prospective observational study.

Results: We report 20 patients with genetically confirmed FA and uniformly reduced

VVOR gain on rapid video-oculography, that is, eye velocity which failed to match head

velocity, resulting in gaze position errors, which were corrected with bursts of saccades

and perceptible as the clinical sign of an impaired VVOR.

Conclusions: This study further elucidates the pathophysiology of the neuro-otological

manifestations of FA. Given the robust and uniform nature of these results, the VVOR is

a planned biomarker for implementation in FA treatment trials.


94

P47

Toward objective clinical diagnosis of cerebellar ataxia

Szmulewicz DJ. 1, 2, 3, Pathirana P.4, Power L.3, Horne M3

1.Balance Disorders and Ataxia Service, Royal Victorian Eye and Ear Hospital,

Melbourne, Australia 2.Cerebellar Ataxia Clinic, Neuroscience Department, AlfredHealth, Melbourne, Australia 3.Florey Institute of Neuroscience and Mental Health, Melbourne, Australia 4.Faculty of Science Engineering and Built Environment, Deakin University, Melbourne,

Australia

Background: One of the most common and medically concerning manifestations of ataxia

(or incoordination) is gait imbalance. Imbalance represents one of the most prevalent

medical complaints globally and in the developed world is as common a presentation as

back pain or headache. With an overall incidence of 5-10%, imbalance effects 40% of

people older than 40 years and the incidence of falls is 25% in those aged 65 and over.

Diagnosis of dizziness or balance disorders is very often challenging, with no single cause

accounting for more than 5–10% of cases. There has also been a lack of ‘tools’ for readily

describing and measuring dysfunction in these systems.

Objectives: This current program of work aims to instrument key aspects of the clinical

examination that are utilized in the assessment of the imbalanced patient.

Materials and methods: Customised inertial measurement units, speech recognition and

visual-kinematic systems have been applied to a set of functional cerebellar domains.

Results: We present objective, stratified data on clinical cerebellar examination metrics.


95

P48

Developing a clinically meaningful instrumented measure of upper limb function in

Friedreich ataxia.

Corben LA1, 2, 3, Tai G1, Szmulewicz D4,5,6, Horne MK6, Pathirana PN7, Delatycki MB1, 2,

3,8 1 Bruce Lefroy Centre for Genetic Health Research, Murdoch Childrens Research

Institute, Parkville, Vic., Australia, 2 School of Psychological Sciences, Monash

University, Clayton, Vic., Australia, 3 Department of Paediatrics, University of

Melbourne, Parkville, Vic., Australia, 4 Balance Disorders and Ataxia Service, Royal

Victorian Eye and Ear Hospital, Melbourne, Australia, 5 Cerebellar Ataxia Clinic,

Neuroscience Department, AlfredHealth, Melbourne, Australia, 6 Florey Institute of

Neuroscience and Mental Health, Melbourne, Australia, 7 School of Engineering, Deakin

University, Geelong, Vic., Australia, 8 Victorian Clinical Genetics Service, Parkville, Vic.,

Australia.

Objectives: Friedreich ataxia (FRDA) has a significant effect on upper limb function which

in turn, compromises independence and quality of life. The most common measure of

upper limb function in FRDA is the Nine Hole Peg Test (9HPT). Increasingly, regulatory

bodies are calling for outcome measures to reflect changes in functional status however

the capacity for the 9HPT to reflect functional capacity is uncertain. The aims of this study

were twofold: 1) to identify the functional upper limb tasks that individuals with FRDA

found most challenging and 2) and use these results, to develop and pilot an instrumented

measure of upper limb function that captures burden of disease and potentially clinically

meaningful change.

Material and methods: We analysed the upper limb component of the Friedreich Ataxia

Impact Scale (FAIS) in 120 individuals with FRDA. In addition, we examined performance

on the Jebsen Taylor Hand Function Test (JHFT) and 9HPT in 73 individuals with FRDA

correlating both measures with clinical parameters of FRDA. Based on this analysis we

developed an instrumented motion capture functional upper limb measure for FRDA.

Results: Intricate tasks such as taking a spoon to the mouth proved to be most

problematic in 88% of participants, significantly correlating with age at disease onset (r=-

0.229, p<0.05), disease duration (r=0.53, p<0.00), the dominant 9HPT (r=0.37, p<0.00)

and all items in the upper limb section of the Friedreich Ataxia Rating Scale (FARS).

Simulated feeding with the dominant hand on the JHFT significantly correlated with

disease duration (rho=0.40, p<0.00) and the 9HPT (rho=0.58, p<0.00).

Conclusion: We have systematically identified a functional task that has provided the

genesis for development of a true measure of upper limb function. This novel

instrumented measure aims to accurately reflect upper limb function in individuals with

FRDA and as such will be of significant utility in future clinical trials.


96


Theme 7- Autism Spectrum Disorder.

P49 / Miss. Chie Morimoto/ Contribution of temporal processing instability with millisecond

accuracy to motor impairments in ASD: an analysis of a synchronized finger-tapping task.

P50 / Dr. Lisa Mapelli/ Autism spectrum disorders and cerebellum: new clues from the IB2 KO

mouse model.

P51 / Dr. Shen Ying/ Ablation of TFR1 in Purkinje cells inhibits mGlu1 trafficking and impairs

motor coordination but not autistic-like behavior.

P52 / Mrs. Mehnosh Toback/ The effects of electromagnetic field (EMF) exposure on cerebellum

development especially regarding autism spectrum disorder (ASD) (Review)

P53 / Mr. Carl O. Olson/ Abnormal molecular properties of cerebellum in Rett Syndrome

patients.


97

P49

Contribution of temporal processing instability with millisecond accuracy to motor

impairments in ASD: an analysis of a synchronized finger-tapping task

Morimoto C. 1, Okamura H. 1

1 Hiroshima University, department of Psychosocial Rehabilitation Institute of

Biomedical & Health Sciences, Hiroshima, Japan

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder, but its underlying

pathogenesis remains largely unknown. Motor impairments are the most common feature

in ASD and some part of motor impairments is assumed to be a result of dysfunctions of

the cerebellum. Therefore, analysis of motor dysfunctions relating to the cerebellum

would be indispensable for evaluation of severity and developmental changes of ASD.

More recent studies suggest that the cerebellum contributes to critically temporal

processing with accuracy down to a millisecond. In the present study, we focused on time

processing with millisecond accuracy to find characteristic marker for motor impairments

of ASD. A synchronized finger-tapping task was employed to evaluate two types of

temporal processing information: (1) phase information and (2) periodic information. The

means and standard deviations (SD) were compared statistically between individuals-

matched along age- sex-with ASD (n=51) and Typical Development (TD) (n=58). To

discriminate age-related change from disease-related change, we compared ASD with

TD across two age groups: 10-14 and 15-19. The averages of temporal processing

parameters showed no significant differences between TD and ASD in both age groups.

However, the SDs of both temporal processing parameters was significantly greater in

ASD than TD for both age groups, with greater variability observed in the younger group.

In addition, receiver operating characteristic (ROC) analyses indicated that altered

temporal processing parameters can play a role in discriminating individuals with ASD

from those without it. These results suggest that altered temporal processing with

millisecond accuracy is a cardinal feature in motor impairments of ASD and useful as the

diagnostic marker for ASD.


98

P50

Autism spectrum disorders and cerebellum: new clues from the IB2 KO mouse model

Mapelli L1, Soda T1,2, Locatelli F1, Botta L3, Goldfarb M4, Prestori F1, D'Angelo E1,5

1 Dept of Brain and Behavioral Sciences, University of Pavia, Pavia, Italy 2 Museo Storico della Fisica e Centro Studi e Ricerche Enrico Fermi, Rome, Italy 3 Dept of Biology and Biotechnology "L. Spallanzani", University of Pavia, Pavia, Italy 4 Dept of Biological Sciences, Hunter College, New York, USA 5 Brain Connectivity Center, C. Mondino National Neurological Institute, Pavia, Italy

Autism spectrum disorders (ASD) are pervasive neurodevelopmental conditions including

familial syndromes with known genetic background. Recent research, despite

heterogeneity in ASD landmarks, in many cases revealed a remarkable involvement of

the cerebellum (Allen, 2006; Hampson and Blatt, 2015). In order to address the role of

cerebellar alterations in ASD pathophysiology, single-neuron and microcircuit properties

were investigated in the IB2 knock-out (KO) mouse model of ASD (Giza et al., 2010),

which corresponds to the Phelan-McDermid syndrome in humans. By exploiting patch-

clamp electrophysiology and voltage-sensitive dye imaging (VSDi), we observed severe

alterations of information processing in the granular layer of IB2 KO mice. The granule

cells showed up to 3.3-times larger NMDA receptor-mediated currents, enhanced intrinsic

excitability, altered excitatory/inhibitory ratio, enhanced long-term potentiation magnitude

and extension. Given these alterations, special attention was devoted to the spatial

organization of granular layer activity in response to inputs, which normally assumes a

center/surround structure. In IB2 KO mice, we found larger centers of excitation with

smaller inhibitory surrounds, revealing a shift from a "Mexican hat" to a "stovepipe hat"

profile (Casanova, 2003, 2006). In aggregate, our results confirm the validity of the IB2

KO mouse as an ASD model and strongly support the cerebellar hypothesis of ASD.

Consistent with observations on neocortical areas, the present data indicate that the

hyper-excitation hypothesis of ASD can be extended to the cerebellum. In detail, the

cerebellar hypothesis predicts that enhanced NMDA receptor functions would cause

hyper-plasticity and derangement of the spatially organized microcircuit activity. The

repercussion of cerebellar alterations inside the whole cerebello-cortical loops (especially

with the prefrontal cortex) opens a new scenario in which multiple circuits concur to ASD

dysfunctions.


99

P51

Ablation of TFR1 in Purkinje cells inhibits mGlu1 trafficking and impairs motor

coordination but not autistic-like behavior

Zhou J.H. 1, Zhou L. 1, Shen Y. 1

1 Department of Neurobiology, Zhejiang University School of Medicine, Hangzhou, P. R.

China.

Group 1 mGlu receptors (mGlu1/5) is critical to synapse formation during development

and participates in long-term potentiation (LTP) and long-term depression (LTD) in the

hippocampus and the cerebellum. mGlu1/5 signaling alterations have been documented

in cognitive impairment, neurodegenerative disorders, and psychiatric diseases.

However, therapeutic strategies for developing mGlu1/5-related agents remain

challenging and the difficult lies in the complex mGlu1/5 signaling. Particularly, the

mechanisms controlling the trafficking of mGlu1/5 are less understood. A recent work

shows that global knockout of Nlgn3 causes an increase in mGlu1 expression in PCs.

Thus, an interesting hypothesis is whether the alteration of mGlu1 signaling in PCs results

in autistic-like behaviors. In the present work, we created a mutant mouse, in which

transferrin receptor 1 (TFR1) was deleted specifically in PCs. The deletion of TFR1 does

not affect the cyto-architecture of PCs, but significantly reduces synaptic expression of

mGlu1 in PCs and inhibits the expression of parallel fiber-LTD. We further found that

TFR1 modulates the trafficking of mGlu1 through Rab proteins. Finally, we demonstrated

that PC ablation of TFR1 impairs motor coordination but does not affect social behaviors.

Together, these results suggest that mGlu1-dependent parallel fiber-PC LTD is

associated with motor learning but not autistic-like behaviors.


100

P52

The effects of electromagnetic field (EMF) exposure on cerebellum development

especially regarding autism spectrum disorder (ASD) (Review)

Toback M.1, Zangeneh K, Marzban A, Akbari M.2, Bergen H.3, Marzban H.3

1Foothills Hospital, 1403, 29 Street N.W., Calgary, Alberta, T2N 2T9, Canada 2Laboratory for Innovations in Microengineering (LiME), Department of Mechanical

Engineering, Center for Biomedical Research, University of Victoria, Victoria, British

Columbia, , V8P 2C5, Canada 3Department of Human Anatomy and Cell Science, Faculty of Health Sciences,

University of Manitoba, Winnipeg, Manitoba, R3E 0J9, Canada,

[email protected]

Because of the increased number of autism spectrum disorder (ASD) patients and

significant increase in electromagnetic field (EMF) exposures over the past few decades,

we hypothesized that there is a link between biological effects of exposure to EMF and

ASD.

ASD is a neurodevelopmental disorder with cerebellum involvement. The cerebellum is

known as a center for motor function and motor skills learning. The cerebellum contains

most of the neurons and synapses in the brain, and therefore it plays a critical role in

emotion and cognitive functions such as language, cognitive processing, and affective

regulation. Deficiencies in cerebellar functions are most likely responsible for ASD

symptoms, patients with abnormalities related to social-cognitive and executive functions

deficits, atypical use of language, and speech difficulties.

Recent studies indicate that exposure to EMF in prenatal life impedes the establishment

and differentiation of neural stem cells and possibly inhibits their transformation into the

neurons during embryonic development. Additionally, EMF induces neural cell death, and

also a major loss of pyramidal cells in the hippocampus of juvenile rats and a substantial

loss of granule cells in the postnatal period. Considerable cerebellar underdevelopment

and a reduced overall number of Purkinje cells, which are significant in ASD, are reported

in response to prenatal exposure to EMF.

Conclusion: This review supports the positive association between ASD and exposure to

EMF, while the increasing number of ASD patients necessitates further research to

explore the adverse effects of EMF during cerebellar development.


101

P53

Abnormal molecular properties of cerebellum in Rett Syndrome patients

Carl Olson. 1, Marc Del Bigio. 2, Victoria Siu. 3, Lee Cyn Ang. 4, Mojgan Rastegar. 1

1 Regenerative Medicine Program, and Department of Biochemistry & Medical Genetics,

Max Rady College of Medicine, Rady Faculty of Health Sciences, University of

Manitoba, MB, Canada, 2 Department of Pathology, Max Rady College of Medicine,

Rady Faculty of Health Sciences, University of Manitoba, MB, Canada, 3 Department of

Biochemistry, Schulich School of Medicine and Dentistry, Western University, ON,

Canada, 4 Department of Pathology, Schulich School of Medicine and Dentistry,

Western University, ON, Canada

Rett Syndrome (RTT) is a severe neurodevelopmental disorder with a frequency of

1:10,000. RTT is associated with neurological symptoms including seizers, anxiety,

breathing problems, and autism. RTT is caused by genetic mutations in an epigenetic

factor that binds to methylated DNA and is called “MeCP2”. Multiple brain regions are

involved in RTT pathobiology, but many animal model studies have focused on

cerebellum. Currently, a gap in knowledge exists on understanding the molecular

abnormalities of human cerebellum in RTT patients.

Objectives: The aim of this study is to uncover the associated molecular interruption of

human cerebellum resulting from common MECP2 mutations. While hundreds of RTT-

associated MECP2 mutations exist, genetic mutations in the 2 functional domains of

MeCP2 (DNA binding domain and transcriptional repression domain) are the cause of

two thirds of RTT cases.

Material and Methods: Human RTT brain tissues are received through donations by family

members from Ontario, Canada, or from NIH NeuroBiobank at the University of Maryland

Brain and Tissue Bank. MeCP2 knockout mice Mecp2tm1.1Bird y/− were obtained from

Jackson labs. Methods of the study include Western blot and immunohistochemical

analysis.

Results and Conclusions: By comprehensive analysis of human RTT cerebellum, we

have identified abnormal characteristics of human cerebellum in the neurons of different

cellular layers of cerebellum. Parallel studies in murine Mecp2-deficient cerebellum

indicate that some of these abnormalities might be specific to human patients. For a better

understanding of the pathobiology of the disease, examination of human RTT brain in

parallel to animal models of RTT is critically important.

This research is supported by International Rett Syndrome Foundation Award#3212,

Ontario Rett Syndrome Association, and Children’s Research Institute of Manitoba. We

are grateful for access to the human brain samples from University of Maryland Brain and

Tissue Bank.


102

Topic 8- Other

P54 / Dr. Takeru Honda/ Novel clinical indexes for evaluating cerebellar internal models

revealed by prism adaptation of human hand-reaching behavior.

P55 / Dr. Franziska Hoche / The Cerebellar Cognitive Affective (CCAS) / Schmahmann

Syndrome Scale

P56 / Dr. Heather Titley/ Clustered Complex Spike Activity Rescues Long-term Depression in

Cerebellar Slices Under Near-Physiological Conditions.

P57 / Dr. Hossein Nahangi/ Effects of adenosine A2a receptor on cerebellar nuclear induced

MDMA toxicity.

P58 / Miss. Kim van Dun/ Polyglot aphasia after left cerebellar stroke.

P59 / Dr. Tadashi Yamazaki/ Large-scale simulation of a cat-scale cerebellar model with one

billion neurons.

P60 /Mr. Shayan Amiri/ Early life stress-induced behavioral abnormalities and mitochondrial

dysfunction in the cerebellum of adult male rats; Effects of pharmacological and non-

pharmacological treatments.

P61 /Prof. Shahram Ejtemaei-Mehr/ The Role of NMDAR/NO Pathway in the Neuroprotective

Effects of Lithium on Cerebellum.

P62 / Mr. Filip Tichanek/ Forced activity subtly mitigates motor and behavioural deficits in

Lurcher mutant mice.

P63 / Dr. Katharina Marie Steiner/ Context-dependency of extinction of conditioned eyeblink

responses in a renewal paradigm in patients with cerebellar lesions.

P64 / Yi-Mei Yang Phosphorylation-specific interaction of potassium channels with Fragile X

mental retardation protein tunes inhibitory neurotransmission


103

P54

Novel clinical indexes for evaluating cerebellar internal models revealed by prism

adaptation of human hand-reaching behavior

Honda T. 1, 3, Matsumura K. 2, Hashimoto Y. 2, Ishikawa K. 2, Mizusawa H. 2, Nagao, S.3

1 Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan 2 Tokyo Med Dent Univ, Tokyo, Japan 3 RIKEN Brain Science Institute, Saitama, Japan

In daily life, we can take any object without viewing the object and our hand attentively

after confirming the object in a short time before the movement. The cerebellar internal

model learning has been assumed to underlie such a feedforward movement control.

Two different internal models have been proposed for cerebellar feedforward movement

control; the inverse model which learns how to move the hand, and the forward model

which learns to move it to where. Which of these two models dose the cerebellum

actually learn has been debated for more than 40 years.

In our study, we proposed theoretically the hybrid cerebellar learning of forward and

inverse models, implementing the long-term depression (LTD) of parallel fiber-Purkinje

cell synapses for learning rule. We applied our model to behavioral data obtained during

prism adaptation from both healthy subjects and patients with cerebellar diseases, and

conducted computer simulations. Our simulations successfully fitted the behaviors of both

healthy subjects and cerebellar patients. We proposed two novel clinical indexes for

explaining the behaviors during prism adaptation: the forward model index (FMI) and the

inverse model index (IMI). Our simulations showed that FMI and IMI represent the abilities

of updating the forward and inverse models, respectively. We compared FMI and IMI

among 10 healthy subjects and 30 (9 MSA-C, 3 MJD, 7 SCA6, 6 SCA31, 4 CCA, and one

SCRA10) cerebellar patients. FMI and IMI distributed in three patterns: ① high FMI and

high IMI in 10 healthy subjects, ② high FMI and low IMI in 21 patients, ③ low FMI and

low IMI in another 9 patients. These observations suggest that the precise motor controls

may be achieved by both the forward and inverse models stored in the cerebellum. The

two novel indexes will help to prepare the protocols of rehabilitations for cerebellar

patients.


104

P55 The Cerebellar Cognitive Affective (CCAS) / Schmahmann Syndrome Scale Franziska Hoche1, Xavier Guell1,2, Mark G. Vangel3, Janet C. Sherman4, Jeremy D. Schmahmann1 1 Ataxia Unit, Cognitive Behavioral Neurology Unit, Laboratory for Neuroanatomy and Cerebellar Neurobiology, Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA., 2 Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA., 3 Martinos Center for Biomedical Imaging, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA., 4 Psychology Assessment Center, Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA Objectives: The cerebellar cognitive affective syndrome (CCAS) is characterized by deficits in executive function, linguistic processing, spatial cognition, and affect regulation (Schmahmann and Sherman, 1998; Schmahmann’s Syndrome - Manto and Mariën, 2015). Diagnosis relies on detailed neuropsychological testing. There is an urgent need for a bedside / office screening test for detection of the CCAS in patients with cerebellar disease. We aimed to: 1) develop a cerebellar cognitive screen to identify CCAS in cerebellar patients, 2) determine whether available brief tests of mental function (Mini Mental State Examination, MMSE; Montreal Cognitive Assessment, MoCA) detect cognitive impairment in cerebellar patients, 3) compare cognitive performance in patients with isolated cerebellar versus those with complex cerebrocerebellar pathology, and 4) detect cognitive deficits that raise red flags about extra-cerebellar pathology. Methods: We tested 77 patients with cerebellar disease – 36 isolated cerebellar degeneration or injury, 41 with complex cerebrocerebellar pathology, and healthy controls on standard and experimental neuropsychological measures. Tests that differentiated patients from controls were used to develop a screening instrument that detects the cardinal elements of the CCAS. This new scale was validated in a separate cohort of 26 cerebellar patients and controls. Results: MMSE and MoCA were not sensitive to cerebellar cognitive impairments. Standard and experimental neuropsychological tests confirmed the defining features of the CCAS, which were present in isolated as well as complex cerebrocerebellar patients. Memory loss was a red flag for non-cerebellar disease. From these results, we derived a 10-item scale that includes semantic fluency, phonemic fluency, category switching, episodic memory, forward and reverse digit span, similarities, cube draw and copy, go/no-go task, and a subjective assessment of affective state. The Scale provides total raw score, cut-offs for each test, and pass-fail criteria that determine Possible, Probable, and Definite CCAS with high sensitivity and selectivity. Conclusion: The Cerebellar Cognitive Affective / Schmahmann Syndrome Scale is useful for the expedited clinical assessment of the CCAS in patients with cerebellar disorders.


105

P56

Clustered Complex Spike Activity Rescues Long-term Depression in Cerebellar Slices

under Near-Physiological Conditions

Titley HK. 1, Kislin M. 2, Wang SS-H. 2, Hansel C. 1

1 University of Chicago, Chicago, Illinois, USA 2 Princeton University, Princeton, New Jersey, USA

Synaptic plasticity at the parallel fiber (PF) to Purkinje cell synapse has been widely

regarded as a correlate for motor learning. In cerebellar slices, long-term potentiation can

be induced by PF stimulation, while long-term depression (LTD) is induced by the

consistent pairing of a complex spike (CS). In previous slice studies, the time interval

required between the PF and climbing fiber (CF) stimulation to obtain LTD has been

suggested to match the time intervals needed between conditioned and unconditioned

stimuli in associative learning tasks (CF delay: 0-150ms; e.g. Wang et al., Nat. Neurosci.

3, 2000). However, recent studies claim that CS-US intervals ≤50ms are inefficient to

drive associate learning (Wetmore et al., J. Neurosci. 34, 2014), and that LTD time

intervals may differ between cerebellar areas (Sruvathan et al., Neuron 92, 2016). To

assess LTD induction parameters under realistic, near-physiological conditions, we

performed whole-cell patch-clamp recordings in mouse slices with GABAergic inhibition

intact, at elevated temperatures (32°C) and using physiological Mg2+ and Ca2+ ASCF

concentrations (1 and 1.2mM, respectively). Under these conditions, a CF pulse paired

with PF stimulation resulted in potentiation at 0, 70, 100, and 200ms PF-CF timing

intervals (total n=13 cells), and no change at 150ms intervals (n=5). In independent in

vivo patch-clamp recordings from Crus I/II Purkinje cells in anesthetized rats (n=7), we

found that complex spikes occurred at a mean firing rate of 1.24 ± 0.08Hz (n=420 CS). In

addition, we found that CS firing can have a clustered pattern with 23% of CSs showing

an inter-CS interval of less than 250ms. In our LTD experiments, we observed that LTD

is rescued (PF-CF interval =150ms) when two complex spikes are evoked with CS-CS

intervals of 150-250ms; n=6). It remains to be determined whether CS cluster stimulation

also rescues LTD at PF-CF timing intervals <150ms.


106

P57

Effects of adenosine A2a receptor on cerebellar nuclear induced MDMA toxicity.

Hossein Nahangi1, Fatemeh Kermanian 2, Mehdi Mehdizadeh3

1Department of Biology & Anatomical Sciences, Shahid Sadoughi University of Medical

Sciences, Yazd, Iran.

BACKGROUND: Adenosine is an endogenous purine nucleoside that has a

neuromodulatory role in the central nervous system. The amphetamine derivative (±)-3,4

methylenedioxymethamphetamine (MDMA or ecstasy) is a synthetic amphetamine

analogue used recreationally to obtain an enhanced affiliated emotional response. MDMA

is a potent monoaminergic neurotoxin with the potential of damage to brain neurons. The

NF-kB family of proteins are ubiquitously expressed and are inducible transcription factors

that regulate the expression of genes involved in disparate processes such as immunity

and ingrowth, development and cell-death regulation. In this study we investigated the

effects of the A2a adenosine receptor (A2a-R) agonist (CGS) and antagonist (SCH) on

NF-kB expression after MDMA administration.

METHODS: Sixty three male Sprague-Dawley rats were injected to MDMA (10 and

20mg/kg) followed by intraperitoneal CGS (0.03 mg/kg) or SCH (0.03mg/kg) injection.

The cerebellum were then removed for cresylviolet staining, Western blot and RT- PCR

analyses. MDMA significantly elevated NF-kB expression. Our results showed that

MDMA increased the number of cerebellar dark neurons.

RESULTS: We observed that administration of CGS following MDMA, significantly

elevated the NF-kB expression both at mRNA and protein levels. By contrast,

administration of the A2a-R antagonist SCH resulted in a decrease in the NF-kB levels.

CONCLUSION: These results indicated that, co-administration of A2a agonist (CGS) can

protect against MDMA neurotoxic effects by increasing NF-kB expression levels;

suggesting a potential application for protection against the neurotoxic effects observed

in MDMA users.


107

P58

Polyglot aphasia after left cerebellar stroke

Van Dun K1, Manto M2, 3, Mariën P1, 4

1 Clinical and Experimental Neurolinguistics, Vrije Universiteit Brussel, Brussels,

Belgium 2 Unité d’Etude du Mouvement, Laboratoire de Neurologie Expérimentale, Université

libre de Bruxelles (ULB), Brussels, Belgium 3 Service des Neurosciences, Université de Mons, Mons, Belgium 4 Department of Neurology and Memory Clinic, ZNA Middelheim General Hospital,

Antwerp, Belgium

We report clinical and fMRI findings in a 72-year-old right-handed man who was submitted

to the hospital after an acute episode of balance problems, vertigo, and vomiting. He

could only speak his maternal language, English, but none of the six languages he

acquired as a late polyglot. MRI revealed an infarction in the vascular territory of the left

posterior inferior cerebellar artery.

In-depth neuropsychological investigations were performed in English one week after

stroke, which revealed no abnormalities except for a deficient score on the Stroop Color

Word test. Formal investigations of language were performed in both English and Dutch.

English was unaffected, but Dutch appeared to be impaired at both the receptive and

expressive level. To investigate lexical-semantic representation, an fMRI naming

paradigm was constructed in English, Dutch, and French, his three best-preserved

languages. fMRI revealed that L1 is lateralized in the language dominant left hemisphere

with a cluster in the left postcentral gyrus and a bilaterally distributed dorsolateral

prefrontal activation (right > left), typically found in multilinguals (response

selection/inhibition). As expected, a larger bilateral network was recruited in the non-

native languages, with stronger activations in the left and right frontotemporal areas, and

cerebellar activations.

From an anatomoclinical point of view, we believe that due to the functional and

anatomical connections between the cerebellum and the prefrontal areas, damage to the

left cerebellar hemisphere could have affected the ability to inhibit the stronger L1,

causing a temporary loss of all non-native languages.


108

P59

Large-scale simulation of a cat-scale cerebellar model with one billion neurons

Yamazaki T. 1, 2, Furusho W. 1

1 The University of Electro-Communications, Graduate School of Informatics and

Engineering, Tokyo, Japan; 2 RIKEN Brain Science Institute, Neuroinformatics Japan Center, Saitama, Japan.

Objectives: Cerebellar corticonuclear microcomplex is a functional unit of the cerebellum,

and the entire cerebellum is thought to be built by copy-and-paste of the microcomplexes.

Previous Theoretical/computational models of the cerebellum have taken only one

microcomplex or multiple independent microcomplexes that do not communicate from

each other into account. However, nearby microcomplexes could share the same parallel

fiber inputs. To understand how multiple microcomplexes communicate and work

together, it is important to build a large-scale model composed of mutually connected

microcomplexes via parallel fibers.

Materials and Methods: We built a large-scale network model of the cerebellum

composed of one billion spiking neurons on a supercomputer, where the network size is

comparable to the whole cerebellum of a cat. We employed several high-performance

computing techniques, such as parallel reduction of calculation of synaptic inputs, and

overlap of calculation and communication over multiple computational nodes. We also

implemented synaptic plasticity including long-term depression and potentiation at

parallel fiber-Purkinje cell synapses. To test the learning capability, we carried out a

computer simulation of gain adaptation of optokinetic response eye movements (OKR),

which is a simple reflex learning task.

Results and Conclusions: In OKR adaptation simulation, our cerebellar model

successfully reproduced the decrease of Purkinje cells' activity due to the long-term

depression of parallel fiber-Purkinje cell synapses caused by climbing fiber inputs, and

increase of the vestibular nuclear neurons' activity resulting in the increase of the

simulated eye movement gain. Furthermore, the computer simulation is performed in

realtime. These results suggest that our cerebellar model would simulate the activity of

the entire cerebellum of a cat during a task, where multiple microcomplexes

communicate each other and work synergistically.


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P60

Early life stress-induced behavioral abnormalities and mitochondrial dysfunction in the

cerebellum of adult male rats; Effects of pharmacological and non-pharmacological

treatments

Amiri S. 1, 2*, Hosseini MJ. 3, Peeri M. 4*

1 current address; Regenerative Medicine Program, Department of Biochemistry and

Medical Genetics, Max Rady College of Medicine, Rady Faculty of Health Sciences,

University of Manitoba, Winnipeg, Manitoba, Canada 2 Department of Pharmacology, School of Medicine, Tehran University of Medical

Sciences, Tehran, Iran 3 Department of Pharmacology and Toxicology, Faculty of Pharmacy, Zanjan University

of Medical Sciences, Zanjan, Tehran 4 Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University,

Tehran, Iran

Increasing lines of evidence indicate that cerebellum is involved in the cognition and

pathophysiology of psychiatric disorders such as depression. It has been well-evident that

experiencing early life adversities is able to negatively affect the brain and behavior in

later life through a variety of mechanisms including mitochondrial dysfunction. Abnormal

mitochondrial function is associated with impairment in energy hemostasis, massive

production of reactive oxygen species, initiation of immune-inflammatory and apoptotic

pathways, and consequently brain damage and dysfunction. Applying maternal

separation stress (MS) to male rats on postnatal day (PND) 2-14, we showed that MS is

able to induce depressive-like behaviors (using forced swimming test, sucrose preference

test and splash test) and cerebellar mitochondrial dysfunction (Increased ROS

production, Decreased ATP and GSH) in adult male rats (PND 60). We also showed that

treating animals with chronic fluoxetine (5 mg/kg/day, i.p), selegiline (1 mg/kg/day, i.p)

and voluntary running wheel exercise during adolescence (PND 30 to PND 60) effectively

attenuated the behavioral deficits and cerebellar mitochondrial dysfunction in adult male

rats. These results suggest that cerebellar mitochondrial dysfunction at least partly is

involved in the pathophysiology of psychiatric disorders such as depression. Also, we

highlighted the importance of adolescence as a period in which treating subjects with

(non) pharmacological therapies is able to mitigate the negative effects of early life stress

on brain and behavior in adulthood.

*Corresponding Authors


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P61

The Role of NMDAR/NO Pathway in the Neuroprotective Effects of Lithium on

Cerebellum

Shahram Ejtemaei Mehr1, 2, Razieh Mohammad Jafari1, 2, Mohammad Hossein

Ghahremani3, Ahmad Reza Dehpour1, 2*

1 Experimental Medicine Research Center, Tehran University of Medical Sciences,

Tehran, Iran, 2 Department of Pharmacology, School of Medicine, Tehran University of

Medical Sciences, Tehran, 3 Department of Toxicology and Pharmacology, Faculty of

Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

Introduction: Cerebellum is a complex structure in the brain, and its abnormal activity is

observed in a variety of brain disorders such as epilepsy. In this regard, cerebellum has

been suggested as the potential target for the treatment of seizure disorders. For

decades, lithium has been used as the main pharmacological agent for the treatment of

bipolar disorder. Evidence suggests that lithium recruits N-Methyl-D-aspartate (NMDA)

receptor/ nitric oxide (NO) pathway to exert its anticonvulsant effects in animal models of

seizures. Since cerebellum is involved in the pathophysiology of seizure disorders, and it

contains high levels of NMDARs, we decided to evaluate whether therapeutic effects of

lithium are associated with modulating of NMDAR/NO pathway in cerebellar granular

cells.

Method: Both in vitro and in vivo studies were conducted. For both model systems, we

used a series of pharmacological treatments with NMDAR antagonist (MK-801), neuronal

NO synthase inhibitor (7-NI), Lithium chloride and the combination of mentioned drugs.

To assess the seizure threshold in mice, pentylentetrazole (PTZ) was administered

intravenously after the 1) chronic treatment of animals with the different doses of lithium

(10, 20 and 30 mg/kg/day, i.p.), 2) lithium + MK-801, 3) lithium + 7-NI and 4) lithium +MK-

801+ 7-NI. For in vitro studies, we used cerebellar granule neurons culture for evaluating

lithium protective effect on cell death in vitro model.

Results: Lithium showed anticonvulsant effects in animals exposed to PTZ. Chronic

lithium treatment before the induction of the clonic seizure significantly increased the

seizure threshold. The serum level of lithium in treated mice were 0.4 mEq/L.

Administration of 7-NI (30 mg/kg, i.p.) had no effect on seizure threshold, but co-

administration of 7-NI before the sub-effective dose of lithium (10 mg/kg, i.p.) decreased

the anticonvulsant effect of lithium significantly. Furthermore, acute injection of MK-801

significantly augmented the anticonvulsant effect of lithium (10 mg/kg, i.p). Also, MK-801

had no significant effect on seizure threshold. Co-administration of low doses of MK-801

and 7-NI with chronic doses of lithium chloride significantly increased the anticonvulsant

effect of lithium.


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Further, by using MTT assay and morphologic examinations, chronic lithium treatment

showed protective effects against glutamate toxicity in cerebellar granule neurons.

Discussion: Our results suggest that NMDAR/NO pathway is involved in anticonvulsant

effects of lithium against PTZ-induced seizures in animals. Also, we showed that lithium

has protective effects against glutamate toxicity in cerebellar granule neurons. Also, more

experiments in this study are in process to show the involvement of NMDAR/NO pathway

in the effects of lithium on cerebellar neurons.


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P62

Forced activity subtly mitigates motor and behavioural deficits in Lurcher mutant mice

Tichanek F. 1, 2, Salomova M. 1, Jelinkova D. 1, 2, Cendelin J. 1, 2

1 Department of Pathological Physiology, Faculty of Medicine in Pilsen, Charles

University, Pilsen, Czech Republic 2 Biomedical Center, Faculty of Medicine in Pilsen, Charles University, Pilsen, Czech

Republic

Physiotherapy represents basic treatment for cerebellar ataxia in humans, modestly

supporting motor skills and thus self-sufficiency of ataxic patients. Analogously to

physiotherapy in humans, forced activity (FA) improves motor deficits in animal models

of cerebellar degeneration via boosting the neuroplasticity. This study aimed to examine

the efficiency of the forced activity as a therapeutic tool for mitigating both the motor and

behavioural dysfunctions in Lurcher mutant mice, a mouse model of cerebellar

degeneration. Once a month, Lurcher mutant mice and their healthy counterparts (starting

at two months of age) were exposed to five day-sessions of either FA training in mowing

treadmill (trained) or to stable treadmill (control). Every month the mice were tested on a

rotarod and the muscle power was measured. After completing four training cycles their

gait was analyzed using CatWalk and DigiGait systems and the behavior was examined

using the open field, elevated plus maze, forced swimming test, novel object recognition,

prepulse inhibition and startle response tests. Only parameters that had been shown to

represent highly sensitive indicator of cerebellar degeneration (i.e. those that were highly

different between Lurcher and Wild-type mice; p<0.001) were used to evaluate effect of

FA. The training significantly improved results in forced swimming test and mitigated

deficits in some gait parameters in Lurcher mice. Although trained Lurcher mice did not

perform better in the rotarod test, compared to controls, they displayed better timecourse

over training months. In conclusion, our preliminary results suggest that FA could partially

mitigate some motor and behavioral deficits in cerebellar mutant mice.

This study was supported by the National Sustainability Program I (NPU I) Nr. LO1503

provided by the Ministry of Education Youth and Sports of the Czech Republic and by the

Charles University Research Fund (project number Q39).


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P63

Context-dependency of extinction of conditioned eyeblink responses in a renewal

paradigm in patients with cerebellar lesions

Steiner K. M. 1, Gisbertz Y. 1, Chang D.-I. 1, Koch B. 1, Uslar E. 1, Claassen J. 1,

Wondzinski E. 2, Goricke S. 3, Siebler M. 2, Timmann D. 1

The cerebellum is known to be involved in both the acquisition and extinction of classical

conditioned eyeblink responses (CRs). Whereas CRs acquired in a certain setting usually

generalize across contexts, CR extinction has been shown to be context-specific, which

is demonstrated by the renewal effect. When CR acquisition takes place in context A and

is unlearned in context B, renewal refers to the reemerging of the CR in context A (ABA-

paradigm). In the present study acquisition, extinction and renewal of classical

conditioned eyeblink responses were tested in an ABA paradigm in patients with focal

cerebellar lesions compared to age-matched healthy controls.

19 patients with subacute cerebellar stroke (12 male, 7 female, mean age 53.1 years,

range 24 to 79 years) and 19 age- and sex-matched healthy controls took part. Standard

delay eyeblink conditioning was performed with a total 140 trials, divided into three

phases (acquisition phase: trial 1-80 in context A, extinction phase: trial 81-110 in context

B, renewal phase: trial 111-140 in context A). Moreover all patients received a 1.5T

structural brain MRI scans that were used for lesion-symptom mapping (LSM).

CR acquisition was not significantly different between cerebellar patients and controls

allowing to draw conclusions on extinction. CR extinction was significantly less in patients

compared to controls. LSM revealed that impaired CR extinction was more likely in

patients with lesions in the lateral lobule VI and Crus I. Controls showed a significant

renewal effect. A significant renewal effect was not present in patients. The present data

provide further evidence that the cerebellum contributes to extinction of conditioned

eyeblink responses in humans. Based on the known cerebello-cerebral connections, Crus

I may contribute to context-related processes in extinction.

Supported by DFG TI 239/10-2.


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P64

Phosphorylation-specific interaction of potassium channels with Fragile X mental

retardation protein tunes inhibitory neurotransmission

Yi-Mei Yang, Jason Arsenault, Alaji Bah, Mickael Krzeminski, Adam Fekete, Laura K

Pacey, Alex Wang, Julie Forman-Kay, David R Hampson and Lu-Yang Wang

Principal neurons encode and project information by varying their firing rates and

patterns that are precisely defined by inhibition from GABAergic interneurons. However,

the molecular basis underlying inhibitory control remains elusive. We find that excessive

presynaptic GABA release from interneurons dramatically attenuates the firing rate of

Purkinje neurons in the cerebellum of Fragile X mental retardation protein (FMRP)

knockout mice. This inhibitory overtone is resulted from increased excitability of the

interneuron terminals where Kv1.2 potassium channels are downregulated in the

absence of FMRP. We further reveal that the N-terminus of FMRP directly binds the C-

terminus of Kv1.2, only when specific serine residues in Kv1.2 are phosphorylated. This

interaction promotes the function of Kv1.2 at the nerve terminal, providing a key

mechanism for dynamic tuning of inhibition; and further understanding of the novel

molecular locus will help develop genetic and pharmacological therapies to rectify

excitation/inhibition imbalance in neuropsychiatric disorders such as Fragile X syndrome

and autism.


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Organizers

Dr. Hassan Marzban (Associate Professor University of Manitoba, Canada) Dr. Mario Manto (Professor, ULB-Erasme, Brussels, Belgium) Program Committee:

Dr. Dan Goldowitz, PC Chair, (Professor, Centre for Molecular Medicine and Therapeutics, Canada) Dr. Timothy Ebner (Professor, University of Minnesota, USA) Dr. Masanobu Kano (Professor, University of Tokyo, Japan) Dr. Kathleen Millen (Professor, Seattle Children's Hospital, USA) Dr. Rachel Sherrard (Professor, Pierre and Marie Curie University, France) Dr. Bing-wen Soong (Professor, National Yang Ming University, Taiwan) Dr. Michisuke Yuzaki (Professor, Keio University School of Medicine, Japan) Dr. Marc Del Bigio (Professor, University of Manitoba, Canada) Dr. Hassan Marzban (Associate Professor University of Manitoba, Canada) Dr. Egidio D'Angelo (Professor, Università di Pavia, Italy) SRCA Executive Committee:

President: Dr. Jean Mariani (Professor, Pierre & Marie Curie University, Paris, France) Vice-President: Marco Molinari (Professor, Translational Research at Fondazione Santa Lucia IRCCS, Rome, Italy) Scientific Chair: Timothy Ebner (Professor, Department of Neuroscience at the University of Minnesota, USA) Secretary: Nori Koibuchi (Professor, Gunma University, Japan) Adjunct Secretary: Esther Becker (Professor, University of Oxford) Treasurer: Jeremy D. Schmahmann (Professor, Massachusetts General Hospital) Editor of The Cerebellum: Mario Manto (Professor, ULB-Erasme, Brussels, Belgium) Local Organizing Committee:

Dr. Mojgan Rastegar, LOC Chair, (Associate Professor, Department of Biochemistry and Medical Genetics) Dr. Michael F. Jackson, LOC Co-Chair, (Assistant Professor, Department of Pharmacology & Therapeutics) Dr. Chris Anderson (Professor and Director, Neuroscience Research Program) Ms. Maryam Rahimi Balaei (PhD student, Department of Human Anatomy and Cell Science) Dr. Hugo Bergen (Associate Professor, Department of Human Anatomy and Cell Science) Dr. Eftekhar Eftekharpour (Assistant Professor, Department of Physiology) Dr. Saeid Ghavami (Assistant Professor, Department of Human Anatomy and Cell Science) Dr. Jean-Eric Ghia (Associate Professor, Departments of Immunology and Internal Medicine section of Gastroenterology) Dr. Ji Hyun Ko (Assistant Professor, Department of Human Anatomy and Cell Science) Dr. Jiming Kong (Professor, Department of Human Anatomy and Cell Science) Mr. Wenyan Li (PhD student, Department of Human Anatomy and Cell Science) Dr. Marc Del Bigio (Professor, University of Manitoba, Canada) Dr. Hassan Marzban (Associate Professor University of Manitoba, Canada)


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Winnipeg Map


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Symposium Sponsors

Department of Human Anatomy and Cell Science

Faculty of Graduate Studies

Peter A. Cattini, Henry G. Friesen Chair in Endocrine & Metabolic Disorders

Department of Biochemistry & Medical Genetics

Department of Pharmacology and Therapeutics

8th - University of Manitoba

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