Characterization of an acrosomal matrix protein in hamster and bovine spermatids and spermatozoa.
Experiments have been carried out characterizing an Mr 22,000 protein present in the acrosomes of hamster and bull spermatozoa. The Mr 22,000 protein is resistant to solubilization in detergent solutions containing high or low salt and has a pI of -5.2. With various lectins, the protein from hamster sperm was shown to be sparingly glycosylated with N-acetylglucosamine, mannose, and galactose while that from the bull demonstrated a slight reactivity for galactose. Using a specific monoclonal antibody (MAB 4/18), the Mr 22,000 polypeptide has been localized exclusively to the acrosomes of mature testicular and epididymal hamster and bovine sperm. Acrosomal components of differentiating bovine and hamster spermatids in tissue sections did not react with the monoclonal antibody, although the protein was present in immunoblots of round spermatids. In bovine sperm, MAB 4/18-staining at the ultrastructural level with immunogold-labeled second antibody was present as a reticulum throughout the acrosomal cap and as punctate aggregates in the equatorial segment. In hamster sperm, MAB 4/18-reactivity was present along the periphery of the acrosome in conjunction with matrix components (M1 and M2), as well as along the inner acrosomal membrane. These observations indicate that the acrosomes of bovine and hamster sperm possess an immunologically related Mr 22,000 protein and suggest that differences in MAB 4/18-staining of spermatids and spermatozoa is a result of epitope modification and/or a change in accessibility of the epitope to the antibody probe during the course of spermiogenesis. Based on its localization and solubility properties, we suggest that the Mr 22,000 protein, in conjunction with other polypeptides, forms a structural framework to maintain acrosomal shape and/or compartmentalize acrosomal contents.
Longo
F J
FJ
Department of Anatomy, University of Iowa, Iowa City 52242.
Cook
S
S
Baillie
R
R
D000177:Acrosome / Q000032:analysis / Q000276:immunology / Q000648:ultrastructure; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D002417:Cattle; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D008297:Male; D008854:Microscopy, Electron; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification*; D013087:Spermatids / Q000032:analysis* / Q000276:immunology / Q000648:ultrastructure; D013094:Spermatozoa / Q000032:analysis* / Q000276:immunology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011506:Proteins