Incorporation and fate of specific sperm plasma membrane components following insemination as revealed by ultrastructural immunocytochemistry.
Studies have been carried out to 1) further characterize sperm specific plasma membrane polypeptides (33 and 35 kDa) that are recognized by a monoclonal antibody previously described (Longo, 1989) and 2) follow the incorporation and dispersal of these proteins within plasmalemmae of monospermic and polyspermic sea urchin (Arbacia punctulata) eggs and oocytes utilizing immunocytochemical methods at the ultrastructural level of observation. Only sperm labeled when incubated with monoclonal antibody to the 33 and 35 kDa proteins followed by colloidal gold-tagged second antibody. Colloidal gold label was observed on the egg plasma membrane immediately after gamete membrane fusion; the amount and extent of label, i.e., the distance from the site of sperm incorporation, increased with time postinsemination. By 20 min postinsemination approximately one hemisphere of the inseminated egg/oocyte was associated with label. The expanding distribution of colloidal gold label on inseminated eggs and oocytes vs. time reflects the free diffusion of 33 and 35 kDa sperm surface proteins among egg/oocyte plasma membrane components. Label was also found in forming endocytotic vesicles, suggesting that sperm plasma membrane proteins may be internalized.
Longo
F J
FJ
Department of Anatomy, University of Iowa, Iowa City 52242.
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000276:immunology; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005306:Fertilization; D006046:Gold; D007150:Immunohistochemistry; D007315:Insemination, Artificial; D008297:Male; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D012617:Sea Urchins; D013094:Spermatozoa / Q000378:metabolism* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D008565:Membrane Proteins; D006046:Gold