Proacrosin/acrosin during guinea pig spermatogenesis.
Enriched populations of guinea pig spermatogenic cells were isolated by sedimentation velocity at unit gravity. Each cell population was analyzed for the presence of members of the proacrosin/acrosin family by enzymography, immunoblotting, and immunofluorescence. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis in gels containing 0.1% gelatin, protease activities with molecular weights of 55,000 (major) and 50,000 (minor) were detected in round spermatid extracts. Condensing spermatid extracts contained protease activities with molecular weights between 55,000 and 50,000. These major protease activities had molecular weights similar to antigens detected by immunoblotting with a monospecific rabbit antiserum directed against purified boar acrosin. Extracts of guinea pig sperm and the soluble acrosomal components released following the acrosome reaction induced with ionophore A23187 contained three major protease activities (Mr 32,000, 34,000, 47,000) but only the 47,000 Mr protease cross-reacted with the antibody. The spermatid and sperm protease activities were inhibited and activated by classical effectors of acrosin activity from other species. Immunofluorescence demonstrated that proacrosin/acrosin was present as early as the Golgi phase of spermiogenesis. In addition, immunoreactivity was confined to the acrosomes in a manner characteristic of each spermatid stage. These results demonstrate that proacrosin/acrosin can be detected in the earliest spermiogenic stages by electrophoretic and immunological techniques and suggest that changes in the molecular weights of proacrosin/acrosin occur as spermatids mature.
D000176:Acrosin / Q000096:biosynthesis*; D000177:Acrosome / Q000032:analysis; D000818:Animals; D003429:Cross Reactions; D004792:Enzyme Precursors / Q000096:biosynthesis*; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D008297:Male; D010447:Peptide Hydrolases / Q000378:metabolism; D012697:Serine Endopeptidases / Q000096:biosynthesis*; D013087:Spermatids / Q000032:analysis; D013091:Spermatogenesis
D004792:Enzyme Precursors; D007461:Iodoacetates; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; C022438:proacrosin; D000176:Acrosin; D019807:Iodoacetic Acid