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Molecular cloning, testicular postnatal expression, and oocyte-activating potential of porcine phospholipase C

¹ Laboratory of Animal Breeding and Reproduction, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan, ² Experimental Farm, Field Science Center (FSC), Hokkaido University, Sapporo 060-0811, Japan and ³ Department of Dairy Science, Rakuno Gakuen University, Bunkyoudai-Midorimachi, Ebetsu, Hokkaido 069-8501, Japan

Correspondence should be addressed to T Watanabe; Email: watanabe{at}anim.agr.hokudai.ac.jp

The molecular mechanism by which sperm triggers Ca²⁺ oscillation, oocyte activation, and early embryonic development has not been clarified. Recently, oocyte activation has been shown to be induced by sperm-specific phospholipase C (PLC). The ability of PLC to induce oocyte activation is highly conserved across vertebrates. In the present study, porcine PLC cDNA was identified and the nucleotide sequence was determined. The expression pattern of porcine PLC mRNA during the period of postnatal testicular development was shown to be similar to that of mouse PLC. PLC mRNA expression in the pig and mouse was detected only in the testes when the elongated spermatids had differentiated, and was detected from day 96 after birth in the pig. Histological examination of porcine testis during the period of postnatal development revealed the presence of spermatozoa from day 110 after birth. These findings suggest that the synthesis of PLC mRNA starts when spermiogenesis is initiated. Microinjection of porcine PLC complementary RNA into porcine oocytes demonstrated that porcine PLC has the ability to trigger repetitive Ca²⁺ transients in porcine oocytes similar to that observed during fertilization. It was also found that porcine PLC cRNA has the potential to induce oocyte activation and initiate embryonic development up to the blastocyst stage.

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