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Molecular characteristics of horse phospholipase C zeta ( PLC ζ)
Author(s) -
Sato Kana,
Wakai Takuya,
Seita Yasunari,
Takizawa Akiko,
Fissore Rafael A.,
Ito Junya,
Kashiwazaki Naomi
Publication year - 2013
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1111/asj.12044
Subject(s) - microinjection , messenger rna , pronucleus , phospholipase c , biology , sperm , microbiology and biotechnology , oocyte activation , chemistry , oocyte , zygote , biochemistry , embryo , gene , signal transduction , genetics , embryogenesis
Abstract A sperm‐specific phospholipase C ( PLC ), PLCzeta ( PLC ζ), is thought to underlie the initiation of calcium ( [Ca 2+ ] i ) oscillations that induce egg activation in mammals. In large domestic species, only bovine, porcine and recently equine PLC ζ have been cloned, and the physiological functions of these molecules have not been fully characterized. Here, we evaluated the physiological functions of equine PLC ζ ( ePLC ζ) in mouse oocytes. ePLC ζ was cloned from testis using RT‐PCR . The expression of ePLC ζ messenger RNA was confirmed in testis but not in other tissues. Microinjection of ePLC ζ complementary RNA ( cRNA ) into mouse oocytes induced long‐lasting [Ca 2+ ] i oscillations, and most of the injected oocytes formed pronuclei ( PN ). The injection of cRNAs encoding horse, mouse, human and cow PLC ζ into mouse oocytes showed that ePLC ζ had the highest [Ca 2+ ] i oscillation‐inducing activity among the species tested. Mutation of D202R , which renders the protein inactive, abrogated the activity of ePLC ζ. The nuclear translocation ability of ePLC ζ was defective when expressed in mouse oocytes. Taken together, our findings show for the first time that ePLC ζ has highest activity of the mammalian species studied to date. Our findings will be useful for the improvement of reproductive technologies in the horse.