Molecular Human Reproduction, Vol. 6, No. 2, 163-169,
February 2000
© 2000 European Society of Human Reproduction and Embryology
Fertilization |
Expression of cadherin adhesion molecules on human gametes
1 Department of Embryology, Sackler School of Medicine and 2 Department of Obstetrics & Gynecology, Rabin Medical Centre, Campus Beilinson, Tel-Aviv University, Israel
Abstract
The presence of cadherins, Ca2+-dependent cell–cell adhesion molecules which may be involved in gamete interaction, was investigated in human gametes. Expression of cadherin molecules was demonstrated using an anti-pan-cadherin antibody and specific antibodies against the three classical cadherins: E- (epithelial), P- (placental) and N- (neural) cadherins. Samples of 48 h old unfertilized oocytes and spermatozoa from in-vitro fertilizing semen samples were lysed and separated by electrophoresis. Localization of cadherins was determined on intact, fixed, permeabilized spermatozoa and oocytes by immunocytochemisty assessed by confocal microscopy. Immunoblotting with the pan-cadherin antibody revealed a single band of ~120 kDa in spermatozoa (whether `fresh', capacitated, or frozen–thawed) and oocyte extracts. Oocytes presented all three classical cadherins with the appropriate molecular weights of 120–130 kDa. In sperm lysate we demonstrated the presence of E-cadherin but not N-cadherin. The anti-P antibody detected a 90 kDa peptide as the only immunoreactive antigen. Following immunocytochemistry of human oocytes all cadherin molecules were allocated predominantly to the plasma membrane with only traces in the cytoplasm. In spermatozoa, several staining patterns were observed with each of the pan-cadherin, N-cadherin and E-cadherin antibodies mostly confined to different head regions. We conclude that cadherin molecules are present on plasma membranes of both human spermatozoa and oocytes and may play a role in the intricate recognition process preceding gamete fusion.
cadherins/gamete fusion/oocytes/spermatozoa
Notes
3 To whom correspondence should be addressed
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