Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print December 15, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.036244
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Submitted September 14, 2004
Returned for revision October 18, 2004
Accepted November 18, 2004

Gamete Biology


Transgenic RNA Interference (RNAi) Reveals Role for Mouse Sperm Phospholipase C{zeta} in Triggering Ca2+ Oscillations During Fertilization

Jason G. Knott , Manabu Kurokawa , Rafael A. Fissore , Richard M. Schultz , and Carmen J. Williams *

* To whom correspondence should be addressed. E-mail: cjwill{at}mail.med.upenn.edu.

Abstract
A sperm-specific phospholipase C, phospholipase C{zeta} (PLC{zeta}), is proposed to be the soluble sperm factor that induces Ca2+ oscillations in mammalian eggs and thus initiates egg activation in vivo. We report here that sperm from transgenic mice expressing short hairpin RNAs (shRNA) targeting PLC{zeta} mRNA have reduced amounts of PLC{zeta} protein. Sperm derived from these transgenic mice trigger patterns of Ca2+ oscillations following fertilization in vitro that terminate prematurely. Consistent with the perturbation in patterns of Ca2+ oscillations is that mating of transgenic founder males to females results in lower rates of egg activation and no transgenic offspring. These data strongly suggest that PLC{zeta} is the physiological trigger of Ca2+ oscillations required for activation of development.

Key words: Gamete Biology • Calcium • Fertilization • Ovum • Sperm


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