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Transgenic RNA Interference Reveals Role for Mouse Sperm Phospholipase C in Triggering Ca²⁺ Oscillations During Fertilization¹

Center for Research on Reproduction and Women's Health,⁴ Department of Biology,⁵ Department of Obstetrics and Gynecology,⁶ University of Pennsylvania, Philadelphia, Pennsylvania 19104 Department of Veterinary and Animal Sciences,⁷ University of Massachusetts, Amherst, Massachusetts 01003

A sperm-specific phospholipase (PL) C, termed PLC, is proposed to be the soluble sperm factor that induces Ca²⁺ oscillations in mammalian eggs and, thus, initiates egg activation in vivo. We report that sperm from transgenic mice expressing short hairpin RNAs targeting PLC mRNA have reduced amounts of PLC 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 the finding 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 is the physiological trigger of Ca²⁺ oscillations required for activation of development.

¹ Supported by grants from the National Institutes of Health (HD22732 to R.M.S. and C.J.W. and HD042498 to R.A.F.) and U.S. Department of Agriculture (02-2078 to R.A.F.). J.K. was supported by training grant T32 HD007305.

² Correspondence: Carmen J. Williams, Center for Research on Reproduction and Women's Health, School of Medicine, University of Pennsylvania, 1313 BRB II/III, 421 Curie Blvd., Philadelphia, Pennsylvania 19104-6080. FAX: 215 573 7627; cjwill{at}mail.med.upenn.edu

³ These authors contributed equally to this work

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