Calcium-dependent and calcium-independent proteinase activities were detected in extracts of rat ventral prostate and its secretion by use of gelatin-containing SDS-PAGE zymography. Ca2+-independent proteinase activities of 22, 26, and 73-79 kDa and Ca2+-dependent activities of 58, 63, and 66 kDa were found in the adult gland. The 26- (most intense activity of gland) and 22-kDa activities were present in secretion and were not expressed in the undifferentiated gland of the 10-day-old animal. The Ca2+-dependent activities were also present in the secretion, where the 63-kDa form was more prominently expressed than the 58- and 66-kDa bands. The Ca2+-dependent and Ca2+-independent proteinase activities both responded to a broad range of pH values in the incubation media. The 73-79-kDa Ca2+-independent activities were sensitive to benzamidine and the Ca2+-dependent activities were inhibited by EDTA and EGTA. Both Ca2+-dependent and Ca2+-independent proteinase activities responded to androgenic manipulations. Castration was followed by the appearance of a 35-kDa Ca2+-independent proteinase (at 2 days) and a 43-kDa Ca2+-dependent proteinase (at 4 days). In the Ca2+-independent proteinase group, the 73-79-kDa activities were increased somewhat and the 22- and 26-kDa activities decreased after castration. The Ca2+-dependent proteinases of 58, 63, and 66 kDa increased in activity with castration, but activity of the 58-kDa form decreased again at 7 days after castration. Treatment of animals upon castration for 4 days with hydrocortisone prevented these changes in proteinase activities whereas treatment with actinomycin D or tranexamic acid did not. Testosterone propionate replacement therapy of rats castrated for 16 days stimulated the activities of the 22- and 26-kDa and 73-79-kDa Ca2+-independent and the 58- and 63-kDa Ca2+-dependent proteinases with 4 days of therapy. The activities of the 35-kDa Ca2+-independent and the 43-kDa Ca2+-dependent proteinases were repressed with 8 days of testosterone treatment. Thus, individual proteinases show differential changes in activity during development and in response to androgenic manipulation: this suggests that in addition to proteinases which are secreted, others may be involved in intracellular functions or in mediating tissue organization changes.
