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Hum. Reprod. Advance Access published online on October 27, 2005
Human Reproduction, doi:10.1093/humrep/dei364
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received April 29, 2005
Revised September 27, 2005
Accepted September 28, 2005

Article

Hyper-osmotic condition enhances protein tyrosine phosphorylation and zona pellucida binding capacity of human sperm

D.Y. Liu 1*, G.N. Clarke 2, and H.W.G. Baker 3

1 Department of Obstetrics and Gynecology, University of Melbourne, Melbourne, Australia
2 Andrology Laboratory, Royal Women’s Hospital, Melbourne, Australia
3 Department of Obstetrics and Gynecology, University of Melbourne, Reproductive Services, Royal Women’s Hospital, Melbourne, Australia; Melbourne IVF, Melbourne, Australia

* To whom correspondence should be addressed.
D.Y. Liu, E-mail: dyl{at}unimelb.edu.au


  Abstract

BACKGROUND: The aim of this study was to determine the effect of culture medium osmolality, in the range known to occur in the male and female reproductive tracts, on human sperm tyrosine phosphorylation and sperm-zona pel-lucida (ZP) interaction in vitro. METHODS: Motile sperm (2x106), selected by swim-up from semen of normo-zoospermic men with normal sperm-ZP binding, were incubated with or without four oocytes in 1 ml human tubal fluid (HTF) medium with different osmolalities (150, 200, 280, 350, 400 mOsm/kg) adjusted by variation of the NaCl concentration. After 2 h incubation, the number of sperm bound to the four ZP was examined, sperm motility and velocities were assessed by Hamilton-Thorn Motility Analyzer (IVOS 10) and sperm tyrosine phosphorylation was assessed by both western immunoblotting and immunofluorescence with an anti-phosphotyrosine monoclonal antibody (PY20). The effect of hyper-osmolality (400 mOsm/kg) on the ZP-induced acrosome reaction (AR) was also determined. RESULTS: Incubation of human sperm in hyper-osmotic medium significantly increased tyrosine phos-phorylation and the number of sperm bound to the ZP. In contrast, hypo-osmotic medium significantly decreased both tyrosine phosphorylation and sperm-ZP binding. Medium with high osmolality (400 mOsm/kg) significantly reduced the ZP-induced AR. Both hypo- and hyper-osmotic media significantly decreased average sperm percentage progressive motility and velocities. CONCLUSION: Incubation of human sperm in hyper-osmotic media was associated with significantly increased tyrosine phosphorylation and ZP-binding ability but severely reduced the ZP-induced AR.

Keywords: capacitation/osmolality/sperm-zona pellucida interaction/tyrosine phosphorylation.
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