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Phospholipase C? rescues failed oocyte activation in a prototype of male factor infertility.

ABSTRACT: OBJECTIVE:To determine the effect of infertility-linked sperm phospholipase C? (PLC?) mutations on their ability to trigger oocyte Ca(2+) oscillations and development, and also to evaluate the potential therapeutic utility of wild-type, recombinant PLC? protein for rescuing failed oocyte activation and embryo development. DESIGN:Test of a novel therapeutic approach to male factor infertility. SETTING:University medical school research laboratory. PATIENT(S):Donated unfertilized human oocytes from follicle reduction. INTERVENTION(S):Microinjection of oocytes with recombinant human PLC? protein or PLC? cRNA and a Ca(2+)-sensitive fluorescent dye. MAIN OUTCOME MEASURE(S):Measurement of the efficacy of mutant and wild-type PLC?-mediated enzyme activity, oocyte Ca(2+) oscillations, activation, and early embryo development. RESULT(S):In contrast to the wild-type protein, mutant forms of human sperm PLC? display aberrant enzyme activity and a total failure to activate unfertilized oocytes. Subsequent microinjection of recombinant human PLC? protein reliably triggers the characteristic pattern of cytoplasmic Ca(2+) oscillations at fertilization, which are required for normal oocyte activation and successful embryo development to the blastocyst stage. CONCLUSION(S):Dysfunctional sperm PLC? cannot trigger oocyte activation and results in male factor infertility, so a potential therapeutic approach is oocyte microinjection of active, wild-type PLC? protein. We have demonstrated that recombinant human PLC? can phenotypically rescue failed activation in oocytes that express dysfunctional PLC?, and that this intervention culminates in efficient blastocyst formation.

SUBMITTER: Nomikos M

PROVIDER: S-EPMC3540263 | biostudies-literature | 2013 Jan

REPOSITORIES: biostudies-literature

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Phospholipase Cζ rescues failed oocyte activation in a prototype of male factor infertility.

Nomikos Michail M Yu Yuansong Y Elgmati Khalil K Theodoridou Maria M Campbell Karen K Vassilakopoulou Vyronia V Zikos Christos C Livaniou Evangelia E Amso Nazar N Nounesis George G Swann Karl K Lai F Anthony FA

Fertility and sterility 20120921 1

<h4>Objective</h4>To determine the effect of infertility-linked sperm phospholipase Cζ (PLCζ) mutations on their ability to trigger oocyte Ca(2+) oscillations and development, and also to evaluate the potential therapeutic utility of wild-type, recombinant PLCζ protein for rescuing failed oocyte activation and embryo development.<h4>Design</h4>Test of a novel therapeutic approach to male factor infertility.<h4>Setting</h4>University medical school research laboratory.<h4>Patient(s)</h4>Donated u ...[more]

PMID: 22999959

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Project description:We recently identified the DPY19L2 gene as the main genetic cause of human globozoospermia (70%) and described that Dpy19l2 knockout (KO) mice faithfully reproduce the human phenotype of globozoospermia making it an excellent model to characterize the molecular physiopathology of globozoospermia. Recent case studies on non-genetically characterized men with globozoospermia showed that phospholipase C, zeta (PLCζ), the sperm factor thought to induce the Ca(2+) oscillations at fertilization, was absent from their sperm, explaining the poor fertilization potential of these spermatozoa. Since 30% of globozoospermic men remain genetically uncharacterized, the absence of PLCζ in DPY19L2 globozoospermic men remains to be formally established. Moreover, the precise localization of PLCζ and the reasons underlying its loss during spermatogenesis in globozoospermic patients are still not understood. Herein, we show that PLCζ is absent, or its presence highly reduced, in human and mouse sperm with DPY19L2-associated globozoospermia. As a consequence, fertilization with sperm from Dpy19l2 KO mice failed to initiate Ca(2+) oscillations and injected oocytes remained arrested at the metaphase II stage, although a few human oocytes injected with DPY19L2-defective sperm showed formation of 2-pronuclei embryos. We report for the first time the subcellular localization of PLCζ in control human sperm, which is along the inner acrosomal membrane and in the perinuclear theca, in the area corresponding to the equatorial region. Because these cellular components are absent in globozoospermic sperm, the loss of PLCζ in globozoospermic sperm is thus consistent and reinforces the role of PLCζ as an oocyte activation factor necessary for oocyte activation. In our companion article, we showed that chromatin compaction during spermiogenesis in Dpy19l2 KO mouse is defective and leads to sperm DNA damage. Together, these defects explain the poor fertilization potential of DPY19L2-globozoospermic sperm and the compromised developmental potential of embryos obtained using sperm from patients with a deletion of the DPY19L2 gene.

Project description:BackgroundAnogenital distance (AGD), the distance between the anus and genitals, is in rodents a well-established marker of early androgen action and has been suggested to be so in humans as well. Thus, a link between human AGD and semen quality and potentially fecundity may exist.ObjectiveThe aim of this study was to assess the association between AGD and male factor infertility and among proven fertile men also time to pregnancy (TTP).Material and methodsAll included men were recruited from and examined at Copenhagen University Hospital - Rigshospitalet, Denmark (N = 388). Men with impaired semen quality were included from infertile couples (N = 128), and men with naturally conceived pregnant partners were invited to participate when their partners had their routine second trimester examination (N = 260). All men underwent a physical examination, completed a questionnaire (including TTP for the fertile men), delivered a semen sample and had a blood sample drawn. The primary exposure was AGDAS measured from the centre of the anus to the posterior base of the scrotum. Associations between AGD and fertility status as well as between AGD and TTP among the fertile men were calculated using multiple logistic regression adjusted for covariates.ResultsAGD did not show a statistically significant association with fertility status. In adjusted logistic regression models, the odds of infertility per 1 cm increase in AGDAS were 1.02 (95% confidence interval [CI]: 0.88; 1.19). Among fertile men, a 1-cm increase in AGDAS was associated with an 8% non-statistically significantly reduced odds of having a longer (>3months) TTP (adjusted odds ratio (OR) = 0.92, 95% CI: 0.76-1.11).ConclusionOur study showed that the clinical application of AGD as a predictor of fertility and fecundity seems to be limited as no associations were observed between AGD and fertility status, nor was the decreased risk of experiencing a longer TTP with longer AGDAS statistically significant.

Dataset Information

Phospholipase C? rescues failed oocyte activation in a prototype of male factor infertility.

Publications

Phospholipase Cζ rescues failed oocyte activation in a prototype of male factor infertility.

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