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Divergent effect of mammalian PLC-zeta in generating Ca2+ oscillations in somatic cells versus eggs

Phillips, Sally Victoria, Yu, Yuansong, Rossbach, Andreas Michael, Nomikos, Michail, Vassilakopoulou, Vyronia, Livaniou, Evangelia, Cumbes, Bevan, Lai, Francis Anthony, George, Christopher H. and Swann, Karl 2011. Divergent effect of mammalian PLC-zeta in generating Ca2+ oscillations in somatic cells versus eggs. Biochemical Journal 438 , pp. 545-533. 10.1042/BJ20101581

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Sperm PLCζ (phospholipase Cζ) is a distinct phosphoinositide-specific PLC isoform that is proposed to be the physiological trigger of egg activation and embryo development at mammalian fertilization. Recombinant PLCζ has the ability to trigger Ca2+ oscillations when expressed in eggs, but it is not known how PLCζ activity is regulated in sperm or eggs. In the present study, we have transfected CHO (Chinese-hamster ovary) cells with PLCζ fused with either YFP (yellow fluorescent protein) or luciferase and found that PLCζ-transfected cells did not display cytoplasmic Ca2+ oscillations any differently from control cells. PLCζ expression was not associated with changes in CHO cell resting Ca2+ levels, nor with a significantly changed Ca2+ response to extracellular ATP compared with control cells transfected with either YFP alone, a catalytically inactive PLCζ or luciferase alone. Sperm extracts containing PLCζ also failed to cause Ca2+ oscillations in CHO cells. Despite these findings, PLCζ-transfected CHO cell extracts exhibited high recombinant protein expression and PLC activity. Furthermore, either PLCζ-transfected CHO cells or derived cell extracts could specifically cause cytoplasmic Ca2+ oscillations when microinjected into mouse eggs. These data suggest that PLCζ-mediated Ca2+ oscillations may require specific factors that are only present within the egg cytoplasm or be inhibited by factors present only in somatic cell lines.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Subjects: R Medicine > RG Gynecology and obstetrics
Uncontrolled Keywords: calcium; Chinese-hamster ovary (CHO) cell; egg; oscillation; phospholipase C (PLC); sperm
Publisher: Portland Press
ISSN: 0264-6021
Last Modified: 27 Mar 2020 02:37

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