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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 ORCID: https://orcid.org/0000-0002-6041-7348, Nomikos, Michail ORCID: https://orcid.org/0000-0001-9761-6327, Vassilakopoulou, Vyronia, Livaniou, Evangelia, Cumbes, Bevan ORCID: https://orcid.org/0000-0001-8529-0040, Lai, Francis Anthony ORCID: https://orcid.org/0000-0003-2852-8547, George, Christopher H. and Swann, Karl ORCID: https://orcid.org/0000-0002-4355-1449 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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Abstract

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: 19 Oct 2022 10:39
URI: https://orca.cardiff.ac.uk/id/eprint/25137

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