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

Abstract

Rises in intracellular Ca2+ concentration ([Ca2+](i)) caused by progesterone, an inducer of the acrosome reaction, or by cyclic nucleotides, possible second messengers, were investigated by Ca2+ imaging of the head of individual mouse sperm. Progesterone induced a [Ca2+](i) rise in a dose- dependent manner (4-40 μM), primarily in the postacrosomal region. For 20- μM progesterone, Ca2+ responses occurred in 42% of sperm, separated into two types: transient type (60% of responding cells; duration, 1-1.5 min; mean amplitude, 335 nM) and prolonged type (40%; >3 min; 730 nM). Prolonged responses required higher doses of progesterone, and their occurrence was enhanced significantly 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 did 8-bromo-cAMP. For 1-mM 8-bromo-cGMP, 90% of cells exhibited transient Ca2+ responses (~1 min; 220 nM), independently of the preincubation time. In Ca2+-free medium, most sperm showed no Ca2+ response to progesterone and 8-bromo-cGMP. Pimozide, a Ca2+ channel blocker, completely blocked prolonged responses and partially inhibited transient responses. These results suggest that progesterone activates at least two distinct Ca2+ influx pathways, with fast or slow inactivation kinetics, and some sperm show both types of response. A cyclic nucleotide- mediated process could participate in the progesterone-induced [Ca2+](i) rise.