Measurement of intracellular calcium levels of human spermatozoa acrosome reacted by electropermeabilization.

Human spermatozoa readily undergo in vitro capacitation and the acrosome reaction when exposed to a single, brief, high-voltage electric pulse in a medium containing calcium. Using the fluorescent calcium indicator, Fura-2, the changes in the intracellular calcium concentration of human sperm following exposure to voltages in the range of 200-2000 V cm-1 were measured. The induction of the acrosome reaction by the electropermeabilization pulse was also monitored. In comparison, changes in calcium uptake during a standard capacitation procedure in which sperm samples were incubated for 7 h were analyzed. At hourly intervals, acrosome reaction induction and intracellular calcium uptake were measured. The results indicate that electropermeabilization rapidly and efficiently produces high populations of acrosome-reacted sperm and a corresponding and associated increase in the intracellular calcium concentration.