Recombinant human phospholipase C zeta 1 induces intracellular calcium oscillations and oocyte activation in mouse and human oocytes

Yoon, Sook Young; Eum, Jin Hee; Lee, Jeoung Eun; Lee, Hoi Chang; Kim, You Shin; Han, Ji Eun; Won, Hyung Jae; Park, Sang Hee; Shim, Sung Han; Lee, Woo Sik; Fissore, Rafael A.; Lee, Dong Ryul; Yoon, Tae Ki

doi:10.1093/humrep/des092

Cited by 87 publications

(63 citation statements)

References 65 publications

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“…We have recently demonstrated that NusA is an effective fusion protein partner for human PLCz, significantly increasing the expression of soluble PLCz protein in E. coli, as well as enhancing the stability of the purified protein over time . Expression of active recombinant human PLCz protein has also been reported by other groups (Kashir et al, 2011;Yoon et al, 2012). Following expression of NusA-PLCz fusion proteins in E. coli and purification by Ni-NTA affinity chromatography, the recombinant proteins were characterized.…”

Section: Enzymatic Characterization Of Human Mouse and Human/mouse Pmentioning

confidence: 80%

Human PLC exhibits superior fertilization potency over mouse PLC in triggering the Ca2+ oscillations required for mammalian oocyte activation

Nomikos

Theodoridou

Elgmati

et al. 2014

Molecular Human Reproduction

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A sperm-specific phospholipase C-zeta (PLCz) is believed to play an essential role in oocyte activation during mammalian fertilization. Sperm PLCz has been shown to trigger a prolonged series of repetitive Ca 2+ transients or oscillations in oocytes that precede activation.This remarkable intracellular Ca 2+ signalling phenomenon is a distinctive characteristic observed during in vitro fertilization by sperm. Previous studies have notably observed an apparent differential ability of PLCz from disparate mammalian species to trigger Ca 2+ oscillations in mouse oocytes. However, the molecular basis and confirmation of the apparent PLCz species difference in activity remains to be provided. In the present study, we provide direct evidence for the superior effectiveness of human PLCz relative to mouse PLCz in generating Ca 2+ oscillations in mouse oocytes. In addition, we have designed and constructed a series of human/mouse PLCz chimeras to enable study of the potential role of discrete PLCz domains in conferring the enhanced Ca 2+ signalling potency of human PLCz. Functional analysis of these human/mouse PLCz domain chimeras suggests a novel role of the EF-hand domain in the species-specific differences in PLCz activity. Our empirical observations are compatible with a basic mathematical model for the Ca 2+ dependence of generating cytoplasmic Ca 2+ oscillations in mammalian oocytes by sperm PLCz.

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Section: Enzymatic Characterization Of Human Mouse and Human/mouse Pmentioning

confidence: 80%

Human PLC exhibits superior fertilization potency over mouse PLC in triggering the Ca2+ oscillations required for mammalian oocyte activation

Nomikos

Theodoridou

Elgmati

et al. 2014

Molecular Human Reproduction

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“…We emphasize the importance of the semiquantitative assay we developed to determine PLC activity. (73,74) to infer PLC activity. Our method allowed us to determine that PIP 2 hydrolysis, induced by ARF6, decreased when DAG was sequestered by the C1 domain of Munc13-1.…”

Section: Discussionmentioning

confidence: 99%

ADP Ribosylation Factor 6 (ARF6) Promotes Acrosomal Exocytosis by Modulating Lipid Turnover and Rab3A Activation

Pelletán

Suhaiman

Vaquer

et al. 2015

Journal of Biological Chemistry

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Background: Sperm acrosomal exocytosis requires GTPases, SNAREs, and a complex lipid signaling. Results: Exocytic stimuli promote ARF6 activation, which accomplishes exocytosis by stimulating PLC and Rab3A. Conclusion: ARF6 induces acrosome calcium efflux and assembles the fusion machinery leading to membrane fusion. Significance: This study explores a novel molecular link between ARF6, PLC, and Rab3A and provides insight into the molecular mechanisms of exocytosis and reproduction.

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“…Several lines of evidence support the role of PLCζ in inducing Ca 2+ oscillation in oocytes; microinjection of either PLCζ complementary RNA (cRNA) or recombinant protein into bovine and murine oocytes induces Ca 2+ oscillations and oocyte activation [8,[12][13][14][15][16]. Moreover, injection of human PLCζ cRNA and recombinant PLCζ protein into human oocytes triggers calcium oscillations and development to the blastocyst stage [9,17]. On the other hand, injection of domain-deletion PLCζ constructs leads to the inability of the protein to generate Ca 2+ oscillations, while reduction of PLCζ expression by RNA interference during Capsule PLCζ seems to be a necessary but not sufficient factor in determining the molecular pathway involved in oocyte activation.…”

Section: Introductionmentioning

confidence: 99%

PLCζ sequence, protein levels, and distribution in human sperm do not correlate with semen characteristics and fertilization rates after ICSI

Ferrer-Vaquer¹,

Barragán²,

Fréour

et al. 2016

J Assist Reprod Genet

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Purpose Sperm-borne PLCζ protein induces Ca 2+ oscillations in the oocyte and is believed to play a major role during oocyte activation. However, its implication in fertilization failure following ICSI is still debated. We analyzed PLCζ gene sequence, protein expression level, and localization in both patients with previous failed fertilization by ICSI and sperm donors with proven fertility in order to assess the association of PLCζ with both sperm characteristics and ability to fertilize. Methods Semen from 15 patients and 13 sperm donors with proven fertility was included in the study. Analysis of the PLCζ gene sequence, protein expression through Western blot, and protein localization by immunofluorescence were performed. Results Two patients with total fertilization failure presented mutations in heterozygosis in the PLCζ gene. Comparison with donor sample sequences displayed comparable SNP allele frequency. Distribution pattern of PLCζ did not vary significantly between donor and patient samples. Levels of PLCζ protein in sperm cells showed an interindividual variability both in patient and donor samples. Several SNPs previously reported in infertile patients were also present in fertile men. ConclusionFailed fertilization occurs even when levels and distribution of PLCζ protein are within normal range. PLCζ seems to be a necessary but not sufficient factor in determining the molecular pathway involved in oocyte activation.

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Recombinant human phospholipase C zeta 1 induces intracellular calcium oscillations and oocyte activation in mouse and human oocytes

Abstract: Repeated [Ca(2+)](i) oscillations and oocyte activation were induced in mouse and human oocytes by microinjection of recombinant hPLCZ1 synthesized in E. Coli. Injection of recombinant protein could thus provide a biological solution for inducing artificial activation of oocytes.

Cited by 87 publications

References 65 publications

Human PLC exhibits superior fertilization potency over mouse PLC in triggering the Ca2+ oscillations required for mammalian oocyte activation

Human PLC exhibits superior fertilization potency over mouse PLC in triggering the Ca2+ oscillations required for mammalian oocyte activation

ADP Ribosylation Factor 6 (ARF6) Promotes Acrosomal Exocytosis by Modulating Lipid Turnover and Rab3A Activation

PLCζ sequence, protein levels, and distribution in human sperm do not correlate with semen characteristics and fertilization rates after ICSI

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