Calcium uptake in human spermatozoa: characterization and mechanisms

Basal 45Ca2+ influx was analyzed in human seminal spermatozoa using a method that allows these highly reactive cells to be easily and safely handled. The uptake was a time-dependent process, with its maximum at 400 s. The kinetics of 45Ca2+ transport was saturating as a function of extracellular Ca2+ concentration with a Km of 429 μM and a V(max) of 1.6 nmol 45Ca2+/mg protein/2.5 min. Depolarizing conditions and the calcium channel blocker verapamil did not affect the uptake; based on this, the presence of operating calcium channels in seminal spermatozoa is excluded. The independence of 45Ca2+ uptake on external concentration of both Na+ and Ca2+ suggests that Na+/Ca2+ exchange does not occur in these cells. The anticalmodulin drug trifluoperazine, the mitochondrial inhibitor antimycin A, and the SH reagents N-ethylmaleimide and mersalyl all inhibited the ion transport. A calmodulin-regulated, energy-requiring, proteinaceous Ca2+ transporter seems to be the main operating mechanism of calcium uptake in human seminal gametes.