期刊
BIOCHEMICAL JOURNAL
卷 438, 期 -, 页码 545-553出版社
PORTLAND PRESS LTD
DOI: 10.1042/BJ20101581
关键词
calcium; Chinese-hamster ovary (CHO) cell; egg; oscillation; phospholipase C (PLC); sperm
资金
- Biotechnology and Biological Sciences Research Council [BB/C513142/1]
- Medical Research Council [G0801676]
- Wellcome Trust [080701/Z/06/Z]
- NCSR Demokritos
- Wellcome Trust [080701/Z/06/Z] Funding Source: Wellcome Trust
- MRC [G0801676] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/C513142/1] Funding Source: researchfish
- British Heart Foundation [FS/09/028/27602] Funding Source: researchfish
- Medical Research Council [G0801676] Funding Source: researchfish
Sperm PLC zeta (phospholipase CO zeta 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 PLC zeta has the ability to trigger Ca2+ oscillations when expressed in eggs, but it is not known how PLC zeta activity is regulated in sperm or eggs. In the present study, we have transfected CHO (Chinese-hamster ovary) cells with PLC zeta fused with either YFP (yellow fluorescent protein) or luciferase and found that PLC zeta-transfected cells did not display cytoplasmic Ca2+ oscillations any differently from control cells. PLC zeta 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 PLC zeta or luciferase alone. Sperm extracts containing PLC zeta also failed to cause Ca2+ oscillations in CHO cells. Despite these findings, PLC zeta-transfected CHO cell extracts exhibited high recombinant protein expression and PLC activity. Furthermore, either PL zeta-transfected CHO cells or derived cell extracts could specifically cause cytoplasmic Ca2+ oscillations when microinjected into mouse eggs. These data suggest that PLC zeta-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.
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