TY - JOUR
T1 - Expression of cadherin adhesion molecules on human gametes
AU - Rufas, O.
AU - Fisch, B.
AU - Ziv, S.
AU - Shalgi, R.
PY - 2000
Y1 - 2000
N2 - 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 immunocytochemistry 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.
AB - 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 immunocytochemistry 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.
KW - Cadherins
KW - Gamete fusion
KW - Oocytes
KW - Spermatozoa
UR - http://www.scopus.com/inward/record.url?scp=0033975083&partnerID=8YFLogxK
U2 - 10.1093/molehr/6.2.163
DO - 10.1093/molehr/6.2.163
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
C2 - 10655458
AN - SCOPUS:0033975083
SN - 1360-9947
VL - 6
SP - 163
EP - 169
JO - Molecular Human Reproduction
JF - Molecular Human Reproduction
IS - 2
ER -