Progesterone Induces Hyperpolarization after a Transient Depolarization Phase in Human Spermatozoa1

Progesterone (P4) induces a membrane depolarization and various ion fluxes (chloride efflux, sodium and calcium influxes), which are required for the human sperm acrosome reaction (AR). By use of the potentiometric fluorescent dye DiSC3(5) and two different technical approaches, the present study aimed to quantify and further analyze P4-induced modifications in membrane potential in capacitated human spermatozoa. Spectrofluorimetric analysis revealed that the mean resting membrane potential of sperm was −58 ± 2 mV (n = 12). When 10 μM P4 was added, the sperm membrane depolarized by ∼+15 mV, partly driven by a Cl− efflux. It subsequently repolarized to reach a significant lower potential than the initial resting potential in two thirds of the tested samples. The flow cytometry analysis showed a heterogeneous resting membrane potential and revealed that the depolarization-hyperpolarization events concerned only subpopulations, between 3% and 40% of the sperm cells according to the samples (n = 7). We hypothesize that P4 has a beneficial effect on the ability of zona pellucida to promote the AR in a sperm subpopulation by increasing the number of hyperpolarized cells presenting a membrane potential that is compatible with the opening of T-type calcium channels by subsequent zona pellucida-induced depolarization.