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Mol. Hum. Reprod. Advance Access published online on July 28, 2005
Molecular Human Reproduction, doi:10.1093/molehr/gah200
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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@oupjournals.org
Received May 29, 2005
Accepted June 6, 2005

Article

Capacitation-dependent concentration of lipid rafts in the apical ridge head area of porcine sperm cells

R.A. van Gestel 1, I.A. Brewis 2, P.R. Ashton 3, J.B. Helms 1, J.F. Brouwers 1, and B.M. Gadella 4*

1 Department of Biochemistry and Cell Biology, Institute of Biomembranes, Utrecht University, Utrecht, The Netherlands
2 The Reproductive Biology and Genetics Group, Division of Medical Sciences, The University of Birmingham, Birmingham; Department of Medical Biochemistry and Immunology and Biostatistics and Bioinformatics Unit, Cardiff University, Cardiff
3 School of Chemical Sciences, The University of Birmingham, Birmingham, UK
4 Department of Biochemistry and Cell Biology, Institute of Biomembranes, Utrecht University, Utrecht, The Netherlands; Department of Farm Animal Health, Graduate School of Animal Health, Utrecht University, Utrecht, the Netherlands

* To whom correspondence should be addressed.
B.M. Gadella, E-mail: b.gadella{at}vet.uu.nl


  Abstract

Lipid architecture of the plasma membrane plays an important role in the capacitation process of the sperm cell. During this process, an increase in membrane fluidity takes place, which coincides with a redistribution of cholesterol to the apical region of the head plasma membrane and subsequently an efflux of cholesterol. Cholesterol is also a major player in the formation of lipid rafts or microdomains in the membrane. Lipid rafts favour specific protein-protein interactions by concentrating certain proteins in these microdomains while excluding others. In this study, we investigated the organization of lipid rafts during in vitro capacitation of boar sperm cells. We report on the presence of the lipid raft-specific proteins caveolin-1 and flotillin-1 in sperm cells. Capacitation induced a change in membrane distribution of these proteins. Lipid analysis on detergent-resistant membranes (DRMs) of sperm cells indicated that capacitation induces a lipid raft concentration rather than a disintegration of lipid rafts, because the total amount of lipid in the DRM fraction remained unaltered. Using a proteomic approach, we identified several major DRM proteins, including proteins involved in capacitation-dependent processes and zona pellucida binding. Our data indicate that sperm raft reorganization may facilitate capacitation-specific signalling events and binding to the zona pellucida.

Keywords: caveolin/cholesterol/cycloolextein/detergent-resistant membrane/flotillin/in vitro fertilization/porcine sperm/proteomics.
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