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Characterization of Intracellular Ca²⁺ Increase in Response to Progesterone and Cyclic Nucleotides in Mouse Spermatozoa¹

^a Department of Physiology, Tokyo Women's Medical University School of Medicine, Tokyo, Japan ^b Department of Obstetrics and Gynecology, Juntendo University School of Medicine, Tokyo, Japan ^c Laboratory of Intracellular Metabolism, Department of Molecular Physiology, National Institute for Physiological Sciences, Okazaki, Japan

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.

FOOTNOTES

First decision: 15 October 1999.

¹ This work was supported by a special research grant for the development of characteristic education from the Japan Private School Promotion Foundation.

² Correspondence: Shunichi Miyazaki, Department of Physiology, Tokyo Women's Medical University, School of Medicine, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. FAX: 81 3 5269 7414; shunm{at}research.twmu.ac.jp

³ Yoshinori Kuwabata passed away on 20 February 2000.

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