Molecular Human Reproduction, Vol. 7, No. 7, 633-640,
July 2001
© 2001 European Society of Human Reproduction and Embryology
Testis and spermatogenesis |
Anti-SLIP1-reactive proteins exist on human spermatozoa and are involved in zona pellucida binding
1 Hormones/Growth/Development Research Group, Loeb Health Research Institute, Ottawa Hospital– Civic Campus, 2 Human In Vitro Fertilization Program, Division of Reproductive Medicine, Department of Obstetrics and Gynecology, Faculty of Medicine, University of Ottawa, 1053 Carling Ave., Ottawa, Ontario K1Y 4E9 and 3 Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, 451 Smyth Rd., Ottawa, Ontario K1H 8M5, Canada
Abstract
Sulpholipid immobilizing protein 1 (SLIP1) is an evolutionarily conserved 68 kDa plasma membrane protein, present selectively in germ cells. We have previously shown that mouse sperm SLIP1 is involved in sperm–zona pellucida (ZP) binding. In this report, we extended our study to the human system. Immunoblotting demonstrated that anti-SLIP1-reactive proteins (mol. wt 68 and 48 kDa) could be extracted from human spermatozoa by an ATP-containing solution, a result that is consistent with observations in other species. Direct immunofluorescence, using Cy3-conjugated anti-SLIP1 IgG, revealed SLIP1 staining over the acrosomal region, with higher intensity at the posterior area. Using the human sperm–ZP binding assay, we demonstrated that pretreatment of human spermatozoa from three donors with anti-SLIP1 IgG revealed lower numbers of zona-bound spermatozoa, as compared to the corresponding control spermatozoa treated with normal rabbit serum IgG. This decrease in zona pellucida binding was not from an antibody-induced decline in sperm motility or an increase in the premature acrosome reaction. The results strongly suggest that anti-SLIP-reactive proteins on human spermatozoa play an important role in ZP binding.
sperm oocyte interaction/sperm surface protein/sulphoglycolipid binding protein/zona pellucida
Notes
4 To whom correspondence should be addressed at Loeb Health Research Institute, 725 Parkdale Ave., Ottawa, Ontario, Canada, K1Y 4E9. E-mail: ntanphaichitr{at}ohri.ca
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