Developmental expression and androgen regulation of 24 kDa secretory proteins by the murine epididymis

Two peptides with a molecular weight of 24 kDa and a P(i) of 8.4-8.8 were found to be synthesized and secreted specifically by the caput epididymis of adult male mice under androgen control. The peptides can interact with spermatozoa. In the present study, the developmental pattern of [35S]-methionine-labelled proteins synthesized by the murine caput epididymis at 10, 20, 30 and 40 days of age were studied using two-dimensional polyacrylamide gel electrophoresis (2D PAGE) and autoradiography. Active synthesis of the 24 kDa proteins was detected in the epididymis from 20 days of age, but secretion of the two peptides was only observed from 30 days of age onwards. To determine whether androgens influenced the active expression of 24 kDa proteins in the developing epididymis, their effect on [35S]-methionine incorporation into proteins was assessed using 2D PAGE. Mice were either castrated, castrated then testosterone injected or simply testosterone injected at 10, 20, 30 or 40 days of age. Androgen control of 24 kDa protein expression was also studies in vitro in epididymal organ culture over a 10-day period, with or without testosterone. Androgens were not involved in the initiation of synthesis of the 24 kDa proteins from days 10 to 20, as shown by in-vivo and in-vivo experiments. However, androgens appeared to be essential for maintaining synthesis and secretion of the proteins from 20 days of age onwards. Administration of excessive testosterone was only able to increase secretion of the 24 kDa proteins in intact male mice aged 40 days.