Inhibition of bovine sperm-oocyte fusion by a monoclonal antibody recognising the TEC-2 epitope on bovine oocytes.
The TEC-2 antigenic determinant is a carbohydrate epitope located on a glycoprotein carrier molecule. In the mouse, this epitope is expressed on the zona pellucida and plasma membrane of the oocyte and is associated with the ZP2 glycoprotein and involved in the secondary sperm receptor mechanism. On the bovine oocyte expression is confined to the plasma membrane. The aim of this study was to determine the role the TEC-2 epitope plays during fertilization in the bovine species using the monoclonal antibody TEC-02. Incubating oocytes with the TEC-02 antibody prior to fertilization inhibited cleavage in a dose-dependent manner-the cleavage rate decreased as the concentration of the antibody increased. Significantly more sperm were bound to oocytes exposed to TEC-02 (12 sperm/oocyte) compared to oocytes that were not incubated with the antibody (4 sperm/oocyte). Oocytes treated with the TEC-02 antibody had a 7.5 +/- 3.2% fusion rate and no cortical granule exocytosis compared with oocytes not exposed to the antibody, with 86.5 +/- 5.8% of sperm-oocyte fusions and release of cortical granules. The block to sperm-oocyte fertilization observed in the pretreated group was overcome using intracytoplasmic sperm injection as the method of fertilization that bypassed the fusion process. Although sperm were binding to the oolemma these results suggest that fusion was not occurring and this may be due to the antibody occupying TEC-2 epitope sites involved in the fusion process. In conclusion, the TEC-2 epitope seems to be involved in sperm-oocyte interaction in the bovine species and appears to be involved specifically during the fusion events of fertilization.
Gougoulidis
T
T
Centre for Early Human Development, Institute of Reproduction and Development, Monash University, Clayton, Victoria, Australia.
Trounson
A
A
Dowsing
A
A
D000818:Animals; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D000954:Antigens, Surface / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000276:immunology* / Q000494:pharmacology; D005260:Female; D005306:Fertilization / Q000502:physiology*; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000276:immunology*; D013084:Sperm-Ovum Interactions / Q000187:drug effects / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D002843:Chromatin; D000939:Epitopes; D008562:Membrane Glycoproteins