Localization of CD9 in pig oocytes and its effects on sperm-egg interaction.
CD9 is a cell surface protein that participates in many cellular processes, such as cell adhesion. Fertilization involves sperm and oocyte interactions including sperm binding to oocytes and sperm-oocyte fusion. Thus CD9 may play an essential role during fertilization in mammals. The present study was conducted to examine whether CD9 is present in porcine gametes and whether it participates in the regulation of sperm-oocyte interactions. The presence of CD9 in ovarian tissues, oocytes and spermatozoa was examined by immunohistochemistry, immunofluorescence and immunoblotting. Sperm binding and penetration of oocytes treated with CD9 antibody were examined by in vitro fertilization. The results showed that CD9 was present on the plasma membrane of oocytes at different developmental stages. A 24 kDa protein was found in oocytes during in vitro maturation by immunoblotting and its quantity was significantly (P < 0.001) increased as oocytes underwent maturation and reached the highest level after the oocytes had been cultured for 44 h. No positive CD9 staining was found in the spermatozoa. Both sperm binding to ooplasma and sperm penetration into oocytes were significantly (P < 0.01) reduced in anti-CD9 antibody-treated oocytes (1.2 +/- 0.2 per oocyte and 16.6% respectively) as compared with oocytes in the controls (2.5 +/- 0.4 per oocyte and 70.3% respectively). These results indicated that CD9 is expressed in pig oocytes during early growth and meiotic maturation and that it participates in sperm-oocyte interactions during fertilization.
Li
Yong-Hai
YH
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China.
Hou
Yi
Y
Ma
Wei
W
Yuan
Jin-Xiang
JX
Zhang
Dong
D
Sun
Qing-Yuan
QY
Wang
Wei-Hua
WH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD / Q000032:analysis* / Q000276:immunology; D002478:Cells, Cultured; D005260:Female; D005307:Fertilization in Vitro; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D009865:Oocytes / Q000737:chemistry*; D009866:Oogenesis / Q000502:physiology*; D010053:Ovary / Q000737:chemistry; D013084:Sperm-Ovum Interactions / Q000276:immunology*; D013094:Spermatozoa / Q000737:chemistry; D060245:Tetraspanin 29; D013997:Time Factors
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D008562:Membrane Glycoproteins; D060245:Tetraspanin 29