Characterization of intracellular Ca2+ increase in response to progesterone and cyclic nucleotides in mouse spermatozoa

Rises in intracellular Ca2+ concentration ([Ca2+](i)) caused by progesteron e, an inducer of the acrosome reaction, or by cyclic nucleotides, possible second messengers, were investigated by Ca2+ imaging of the head of individ ual mouse sperm. Progesterone induced a [Ca2+](i) rise in a dose-dependent manner (4-40 mu M), primarily in the postacrosomal region. For 20-mu M prog esterone, Ca2+ responses occurred in 42% of sperm, separated into two types : transient type (60% of responding cells; duration, 1-1.5 min; mean amplit ude, 335 nM) and prolonged type (40%; >3 min; 730 nM). Prolonged responses required higher doses of progesterone, and their occurrence was enhanced si gnificantly by preincubation for 2-4 h as compared with transient responses . 8-Bromo-cGMP (0.3-3 mM) induced a [Ca2+](i) rise more effectively than di d 8-bromo-cAMP. For 1-mM 8-bromo-cGMP, 90% of cells exhibited transient Ca2 + responses (similar to 1 min; 220 nM), independently of the preincubation time. In Ca2+-free medium, most sperm showed no Ca2+ response to progestero ne and 8-bromo-cGMP, Pimozide, a Ca2+ channel blocker, completely blocked p rolonged responses and partially inhibited transient responses. These resul ts suggest that progesterone activates at least two distinct Ca2+ influx pa thways, with fast or slow inactivation kinetics, and some sperm show both t ypes of response. A cyclic nucleotide-mediated process could participate in the progesterone-induced [Ca2+](i) rise.