Absence of active protein kinase C in ram spermatozoa

At fertilization, mammalian spermatozoa undergo a Ca2+-dependent exocytotic event known as the acrosome reaction. As protein kinase C (PKc) has been implicated in exocytosis in some other cell systems, we have searched for PKc in ram spermatozoa. We have found that: (a) no changes in protein phosphorylation pattern could be induced in the intact cells by phorbol dibutyrate (PDBu), a compound which binds to and stimulates PKc; (b) no changes in protein phosphorylation pattern could be detected during the course of the Ca2+/ionophore-induced acrosome reaction (when greater than 95% of the cells underwent exocytosis); (c) there was no effect of PDBu on the exocytotic response to various Ca2+ and ionophore levels; (d) no specific PDBu binding could be detected in the cells (this binding is considered to be indicative of the presence of active PKc). We conclude that potentially active PKc is not present in ram spermatozoa.