Inhibition of the human sperm acrosome reaction by proteinase inhibitors

Abstract

The analogue of the second messenger cAMP, dibutyryl cAMP (dbcAMP), was shown to induce the human sperm acrosome reaction to the same extent as calcium ionophore A23187, providing preliminary evidence for the involvement of the adenylate cyclase system in the acrosome reaction (AR) of human spermatozoa. Using the human synchronous acrosome reaction system, proteinase inhibitors were tested for their effect on the dbcAMP-induced human sperm acrosome reaction. The proteinase inhibitor 4'-acetamidophenyl 4-guanidinobenzoate (AGB), an inhibitor of proacrosin activation and of acrosin, when added at either the onset of incubation or to capacitated spermatozoa, 5 min prior to stimulation by dbcAMP, significantly (P less than 0.01) inhibited the acrosome reaction at final concentrations of 1 x 10(-4) M to 1 x 10(-6) M in comparison to dbcAMP treatment alone. At concentrations less than 1 x 10(-6) M, no significant inhibitory effect was seen. Similarly, para-aminobenzamidine (pAB), also an inhibitor of proacrosin activation and of acrosin, significantly (P less than 0.01) inhibited the dbcAMP-induced acrosome reaction at final concentrations of 1 x 10(-4) M to 1 x 10(-6) M when added at either the onset of incubation or to capacitated spermatozoa, 5 min prior to stimulation by dbcAMP, in comparison to stimulation by dbcAMP alone. However, at concentrations less than 1 x 10(-6) M, no significant (P greater than 0.05) inhibitory effect was seen. These results indicate that a serine proteinase, most likely acrosin, has a role in the human sperm acrosome reaction and suggest that the enzyme functions after the involvement of the adenylate cyclase system.