Research group of Functional Genomics and Proteomics

• - 9 postdocs• - 11 PhD students• - 1 IOF manager• - 4 technicians

http://bio.kuleuven.be/df/ls/people

Research group of Functional Genomics and Proteomics

Human brain ~100 billion

neurons

Caenorhabditis elegans 302 neurons

Neuropeptides are key regulators of animal physiology and behaviour through activation of GPCRs.

Conserved mechanisms: reproduction, locomotion, learning, digestion, ageing…

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Neuropeptides, neuronal communication and neuroplasticity

Big ideas from small brains

• Learning, memory, and ageing• Cancer• Feeding behavior• Reproduction• Sleeping disease

Main research topics:

Characterization of novel neuropeptide-mediated signaling systems in C. elegans

Screening for GPCR ligand

Expression analysis

Behavioral assays

e.g. locomotion RNAi knockdown

Neuropeptides are key regulators of animal physiology (feeding, reproduction) through binding of GPCRs.

C. elegans: neurobiological model

Contact:Contact: Isabel.beets@bio.kuleuven.beIsabel.beets@bio.kuleuven.be

Caenorhabditis elegans advantages:302 neuronen, toolbox for genetic manipulationsthousands of mutant strains available

The locomotive behaviour of the worm can be manipulated by light by making transgene worms in which specific neurons express light-sensitive ion channels, Channelrhodopsine-2 (ChR2, depolarisation, activation) and Halorhodopsine.

Photoactivation of defined neurons in the head let the worms crawl in the shape of a triangleImage modified from Stirman et al. (2011) Nature Methods 8: 153-158

Understanding experience-based learning behaviour: The unraveling of neuronal circuits by innovative optogenetic tools.

Aim: can we control learning behaviour using optogenetics?

A forward mutagenesis screen to study the neuroendocrine regulatory system of reproduction in C. elegans

Mutagenesis

GFP expression in intestine

Fluorescent worms

(vit-2::GFP)COPAS worm sorter

Non fluorescent worms

Gene mappingOR

Whole genome sequencing

Target phenotypingDefects with respect to

reproductionE.g. egg-laying defect

Proteomics and peptidomics of long-lived Proteomics and peptidomics of long-lived C. elegansC. elegans

Techniques

Many life-extending interventions available in C. elegans→ insulin pathway mutations, dietary restriction,…

Studying long-lived worms to help us combat aging-associated diseases→ interventions that increase lifespan also decrease incidence of cancer, neurodegenerative diseases,…

PeptidomicsDifferential labeling of peptides Liquid chromatographyMass spectrometry

ProteomicsGel electrophoresis (2D-DIGE)Mass spectrometry

What are the effectors What are the effectors that control aging?that control aging?

How do the endocrine How do the endocrine systems influence aging?systems influence aging?

Contact:Contact: Wouter.dehaes@bio.kuleuven.beWouter.dehaes@bio.kuleuven.be

GnRHR achtige receptoren bij C. elegans: Reproductie of energiehuishouding?

CONTACT:Lotte.frooninckx@bio.kuleuven.beResearch group of Functional Genomics and Proteomics

VERTEBRATENReceptor wordt geactiveerd door het gonadotropin-releasing hormoon (GnRH) dat de reproductie reguleert.

INSECTENReceptor wordt geactiveerd

door adipokinetisch hormoon (AKH)

dat de energiehuishouding reguleert. C. elegans

8 homologen: gnrr-1 tot gnrr-8Waar komen deze genen tot

expressie?GFP-lokalisatie, PCR, microscopie

Wat is de invloed van een deletie in deze genen: fenotypering, microscopie,microfluidica

Wat zijn de liganden?‘ontwezen receptoren’, celculturen, high-throuput screenen

Neuropeptide hormones and their receptors: a suitable target to combat tsetse flies?

Tsetse flies: vectors for parasites that cause sleeping disease in human and nagana in cattle. There is an urgent call for the development of new species-specific insecticides.G protein-coupled receptors are promising targets for the development of insecticides based on peptidomimetica.

The tsetse fly genome has been sequenced recently and a large number of putative neuropeptides and GPCRs are predicted. The goal is to deorphanize some GPCRs, study their tissue distribution and investigate the binding characteristics of the neuropeptides.

Techniques: - Molecular biology RNA/DNA purification, PCR, sequencing, Reverse pharmacology, Quantitative real-time PCR, Structure-activity studies

Promotor: Prof. Dr. L. Schoofs (http://bio.kuleuven.be/df/LS/)Mentor: Jelle Caers (contact: Jelle.Caers@bio.kuleuven.be)

2D-DIGE2D-DIGE

Differential Differential proteomicsproteomics

Microarray-Microarray-analysisanalysis

TranscTranscript-ript-

omicsomicsMass Mass

spectrometryspectrometry

PeptidomicsPeptidomics

RNA RNA interferenceinterference

Gene SilencingGene Silencing

gregarious solitary

Phenotypic plasticity of physiology and behaviour

Next Next generation generation sequencingsequencing

Elucidation of locust phase polyphenism with gel based proteomics.

2 phenotypes

1 Genome

Epigenetic component?

DNAMethyl-Cytidine

Differential DNA methylation?

Differential PROTEOME?

Workflow:

Tissue dissection

Protein extraction

Differential analysis: 2D-DIGE

Protein identification- MALDI TOF/TOF mass spectrometry- Bioinformatics de novo protein identification

Brain

Bart.Boerjan@bio.kuleuven.be

More info?

Towards a novel insect pest control strategy

More info: Bart.Boerjan@bio.kuleuven.be

Creativity

Bioinformatics

Genetic engineering

confidential

Extensive training in:-proteomics-peptidomics-cell culture-scientific reasoning(SDS-PAGE, 2D-DIGE, mass spectrometry)

Royality in honeybees-the making of a queen

Uli.Ernst@bio.kuleuven.be

Royal jelly, active (protein)

compound: royalactin, 57kDa

royalactin: activates epidermal growth factor

receptor (Egfr) in the fat body

-Epigenetic changes-High juvenile hormone titer-...

...but: how can a 57kDa protein reach the fat body

via the gut???

Annona Cherimoya & cancer

Septic shock

Lethal dose

Sub-Lethal dose

Protected against

Identification of protective factor