L5 huszar slide selection
Transcript of L5 huszar slide selection
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Objective biochemical markers of sperm maturity and function
(Relationship between sperm maturity, fertility and membrane properties)
Sperm-hemizona
complex
Only the clear headed sperm, which have completed cytoplasmic extrusion and
plasma membrane remodeling (proven), were able to bind to the zona pellucida.
Huszar et al., Mol Reprod Dev 1993; Fertil Steril,1994; BOR, 1997
CK immunostaining
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Huszar et al. BOR, 2000
Spermiogenesis: Sperm with Normal
and Diminished Development Mature
Development Development
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DNA Nick Translation Aniline blue CK Sperm field pairs treated by various biomarkers (semen and HA-selected sperm)
semen
HA selected
CK-immuno Persistent histones DNA chain fragmentation
Huszar et al., 2007
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Acridin orange treated spermatozoa in semen and in HA-selected sperm
fraction (red sperm: DNA fragmentation, green sperm: high DNA integrity)
Yagci and
HUSZAR, 2010
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PICSI SPERM SELECTION
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(Jakab et al., F+S, 2005)
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Clinical Loss Rate
0.067 0.011
< 0.001
KCWorrilow et al, 2013
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Sperm Selection Methods
Except HA-selected sperm, little research on sperm attributes, such
as chromatin, DNA integrity, and chromosomal aneuploidies.
Hyaluronic acid-mediated sperm selection
Density gradient
IMSI
Microfluidics
Electrophoretic separation
Birefrigence (polarized light)
Raman spectroscopy (single sperm too, recent
paper-ZP-bound sperm)
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PROPERTIES OF HYALURONIC ACID-BOUND SPERM
EXCELLENT SPERM VIABILITY
NO SURPLUS CYTOPLASMIC CONTENT
KRUGER NORMAL MORPHOLOGY
NO PERSISTENT HISTONES, thus normal histone-protamin progression (DNA PACKING)
HIGH DNA CHAIN INTEGRITY, NO FRAGMENTATION (THREE STUDIES shown)
NO APOPTOTIC PROCESSES
NORMAL FRQUENCY OF CHROMOSOMAL ANEUPLOIDIES
CURRENT ONGOING: RNA STUDIES, EPIGENETIC STUDIES
(Wish list of the IVF-ICSI Profession)
In all aspects studied zona pellucida and HA-bound sperm are identical
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Destaining
FISH
Nick Translation
Caspase-3 Immunostaining
Localization
+ Imaging
Re-localization
+ Imaging
Preparation, Fixation
Aniline Blue staining
of persistent histones
SATI, L., et al., RBMonline, 2008
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Aniline blue staining pattern and DNA fragmentation
(in situ nick translation)