Superoxide anion production by human spermatozoa as a part of the ionophore-induced acrosome reaction process

Summary The involvement of superoxide anion (O2°‐) in human sperm capacitation and/or acrosome reaction was investigated. Addition of superoxide dismutase (SOD) to the medium at the beginning of the capacitation process or 15 min before induction of the acrosome reaction, decreased the level of ionophore‐induced acrosome reaction. Hyperactivation was unaffected by the presence of SOD during the capacitation process. Addition of calcium ionophore to the sperm suspension increased production of O2°‐ by the spermatozoa by four to five‐fold and induced the acrosome reaction. In the presence of SOD, superoxide anion could not be detected in the medium and the rate of induced‐acrosome reaction was decreased gready. The presence of an inhibitor of protein kinase C inhibited the production of O2°‐ in the medium and reduced the induced‐acrosome reaction. The production of O2°‐ and the acrosome reaction were also increased by exposure of spermatozoa to 12‐myristate 13‐acetate phorbol ester, a specific activator of protein kinase C. While the level of spontaneous acrosome reaction was not increased by the direct addition of O2°‐ to the medium, its presence induced the release of unesterified fatty acids from membrane phospholipids. These findings suggest that the production of O2°‐ by spermatozoa could be involved in the ionophore‐induced acrosome reaction, possibly through the de‐esterification of membrane phospholipids. However, this production of superoxide anion is not sufficient on its own to induce the acrosome reaction.