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A CHEMICAL PROCEDURE FOR EXTRACTION OF THE ACROSOMES OF MAMMALIAN SPERMATOZOA

Summary.: Rabbit, human and bull spermatozoa treated with 9·0 mM-barbituric acid reveal elevation and rupture of the acrosomal membrane, as well as partial extraction of acrosomal material. Other sperm membranes do not appear to be affected. A subsequent detergent extraction (Triton X-100) solubilizes the remaining acrosomal material, membranes and associated structures. The action of barbituric acid on the acrosomes of the three types of spermatozoa is immediate, efficient and effective in low temperature ranges. The amount of protein extracted from spermatozoa was characteristic for each species examined. Major portions of protein were extracted with barbituric acid; the remaining acrosomal protein was obtained with the detergent. Enzymes identified in the extracts include acid phosphatase, β-glucuronidase, hyaluronidase and proteases, with acid, neutral and alkaline pH optima. The proteases were unique for each species.