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Published 14 April 2003. doi:10.1083/jcb.200210057
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© The Rockefeller University Press, 0021-9525/2003/4/79 $5.00
The Journal of Cell Biology, Volume 161, Number 1, 79-88

Article

Phospholipase C{delta}4 is required for Ca2+ mobilization essential for acrosome reaction in sperm



Kiyoko Fukami1,5, Manabu Yoshida2,6, Takafumi Inoue2,6, Manabu Kurokawa4, Rafael A. Fissore4, Nobuaki Yoshida3, Katsuhiko Mikoshiba2,6,7 and Tadaomi Takenawa1,5

1 Division of Biochemistry, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
2 Division of Molecular Neurobiology, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
3 Division of Gene Expression and Regulation, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
4 Department of Veterinary and Animal Sciences, Paige Laboratory, University of Massachusetts, Amherst, MA 01003
5 Center for Renewable Energy and Sustainable Technology, International Cooperative Research Project, Japan Science and Technology Corporation, Tokyo 108-8639, Japan
6 Calcium Oscillation Project, International Cooperative Research Project, Japan Science and Technology Corporation, Tokyo 108-8639, Japan
7 RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan

Address correspondence to Kiyoko Fukami, Division of Biochemistry, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Tel.: 81-3-5449-5507. Fax: 81-3-5449-5417. E-mail: kfukami{at}ims.u-tokyo.ac.jp

Zona pellucida (ZP)–induced acrosome reaction in sperm is a required step for mammalian fertilization. However, the precise mechanism of the acrosome reaction remains unclear. We previously reported that PLC{delta}4 is involved in the ZP-induced acrosome reaction in mouse sperm. Here we have monitored Ca2+ responses in single sperm, and we report that the [Ca2+]i increase in response to ZP, which is essential for driving the acrosome reaction in vivo, is absent in PLC{delta}4-/- sperm. Progesterone, another physiological inducer of the acrosome reaction, failed to induce sustained [Ca2+]i increases in PLC{delta}4-/- sperm, and consequently the acrosome reaction was partially inhibited. In addition, we observed oscillatory [Ca2+]i increases in wild-type sperm in response to these acrosome inducers. Calcium imaging studies revealed that the [Ca2+]i increases induced by exposure to ZP and progesterone started at different sites within the sperm head, indicating that these agonists induce the acrosome reaction via different Ca2+ mechanisms. Furthermore, store-operated channel (SOC) activity was severely impaired in PLC{delta}4-/- sperm. These results indicate that PLC{delta}4 is an important enzyme for intracellular [Ca2+]i mobilization in the ZP-induced acrosome reaction and for sustained [Ca2+]i increases through SOC induced by ZP and progesterone in sperm.

Key Words: phospholipase C; acrosome reaction; zona pellucida; calcium; sperm

The online version of this article includes supplemental material.

* Abbreviations used in this paper: IP3, inositol 1,4,5-trisphosphate; IP3R, IP3 receptor; SBTI, soybean trypsin inhibitor; SOC, store-operated channel; ZP, zona pellucida.


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