4.5 Article

Divergent effect of mammalian PLCζ in generating Ca2+ oscillations in somatic cells compared with eggs

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

  1. Biotechnology and Biological Sciences Research Council [BB/C513142/1]
  2. Medical Research Council [G0801676]
  3. Wellcome Trust [080701/Z/06/Z]
  4. NCSR Demokritos
  5. Wellcome Trust [080701/Z/06/Z] Funding Source: Wellcome Trust
  6. MRC [G0801676] Funding Source: UKRI
  7. Biotechnology and Biological Sciences Research Council [BB/C513142/1] Funding Source: researchfish
  8. British Heart Foundation [FS/09/028/27602] Funding Source: researchfish
  9. Medical Research Council [G0801676] Funding Source: researchfish

向作者/读者索取更多资源

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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