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Cited by 157Reproductive Biology and FertilitySperm and Testicular FunctionReproductive Health and Technologies

PLCζ is the physiological trigger of the Ca2+ oscillations that induce embryogenesis in mammals but conception can occur in its absence

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Hachem, Alaa, Godwin, Jonathan, Ruas, Margarida, Lee, Hoi Chang, Ferrer Buitrago, Minerva, Ardestani, Goli, Bassett, Andrew, Fox, Sebastian +5 more·2017·Development
DOI 10.1242/dev.150227PMC PMC5592814
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Abstract

Activation of the egg by the sperm is the first, vital stage of embryogenesis. The sperm protein PLCζ has been proposed as the physiological agent that triggers the Ca2+ oscillations that normally initiate embryogenesis. Consistent with this, recombinant PLCζ induces Ca2+ oscillations in eggs and debilitating mutations in the PLCZ1 gene are associated with infertility in men. However, there has been no evidence that knockout of the gene encoding PLCζ abolishes the ability of sperm to induce Ca2+ oscillations in eggs. Here, we show that sperm derived from Plcz1–/– male mice fail to trigger Ca2+ oscillations in eggs, cause polyspermy and thus demonstrate that PLCζ is the physiological trigger of these Ca2+ oscillations. Remarkably, some eggs fertilized by PLCζ-null sperm can develop, albeit at greatly reduced efficiency, and after a significant time-delay. In addition, Plcz1–/– males are subfertile but not sterile, suggesting that in the absence of PLCζ, spontaneous egg activation can eventually occur via an alternative route. This is the first demonstration that in vivo fertilization without the normal physiological trigger of egg activation can result in offspring. PLCζ-null sperm now make it possible to resolve long-standing questions in fertilization biology, and to test the efficacy and safety of procedures used to treat human infertility.

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  • PLCζ1 is sperm-specific and is the physiological trigger responsible for generating IP3-mediated Ca2+ oscillations that induce oocyte activation during mammalian fertilisation.

    “This PLC is sperm-specific and is the physiological trigger responsible for generating I(1,4,5)P3-mediated Ca2+ oscillations that induces oocyte activation during mammalian fertilisation [43,44].”

    From Phosphatidylinositol(4,5)bisphosphate: diverse functions at the plasma membrane (2020) · cites this paper as [43]

    The Introduction identifies PLCζ as a sperm protein, and the Results section 'Loss of PLCζ abolishes the ability of the sperm to trigger Ca2+ oscillations in the egg' calls it the physiological trigger of oscillations responsible for mammalian egg activation (page unavailable). Our copy does not establish that these oscillations are IP3-mediated; the inositol 1,4,5-trisphosphate mention found is in a reference title, not the paper’s account of this mechanism. (LLM, conf=0.87)

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Research works printed but not linked: 46

  1. Aarabi, M., Yu, Y., Xu, W., Tse, M. Y., Pang, S. C., Yi, Y.-J., Sutovsky, P. and Oko, R. (2012). The testicular and epididymal expression profile of PLCzeta in mouse and human does not support its role as a sperm-borne oocyte activating factor. PLoS ONE 7, e33496. · DOI
  2. Aarabi, M., Balakier, H., Bashar, S., Moskovtsev, S. I., Sutovsky, P., Librach, C. L. and Oko, R. (2014). Sperm-derived WW domain-binding protein, PAWP, elicits calcium oscillations and oocyte activation in humans and mice. FASEB J. 28, 4434-4440. · DOI
  3. Bassett, A. and Liu, J.-L. (2014). CRISPR/Cas9 mediated genome engineering in Drosophila. Methods 69, 128-136. · DOI
  4. Bernhardt, M. L., Lowther, K. M., Padilla-Banks, E., McDonough, C. E., Lee, K. N., Evsikov, A. V., Uliasz, T. F., Chidiac, P., Williams, C. J. and Mehlmann, L. M. (2015). Regulator of G-protein signaling 2 (RGS2) suppresses premature calcium release in mouse eggs. Development 142, 2633-2640. · DOI
  5. Bianchi, E., Doe, B., Goulding, D. and Wright, G. J. (2014). Juno is the egg Izumo receptor and is essential for mammalian fertilization. Nature 508, 483-487. · DOI
  6. Cheng, Y., Zhong, Z. and Latham, K. E. (2012). Strain-specific spontaneous activation during mouse oocyte maturation. Fertil. Steril. 98, 200-206. · DOI
  7. Doudna, J. A. and Charpentier, E. (2014). Genome editing. The new frontier of genome engineering with CRISPR-Cas9. Science 346, 1258096. · DOI · Paper page
  8. Ducibella, T., Huneau, D., Angelichio, E., Xu, Z., Schultz, R. M., Kopf, G. S., Fissore, R., Madoux, S. and Ozil, J.-P. (2002). Egg-to-embryo transition is driven by differential responses to Ca(2+) oscillation number. Dev. Biol. 250, 280-291. · DOI
  9. Escoffier, J., Lee, H. C., Yassine, S., Zouari, R., Martinez, G., Karaouzeǹe, T., Coutton, C., Kherraf, Z.-E., Halouani, L., Triki, C. et al. (2016). Homozygous mutation of PLCZ1 leads to defective human oocyte activation and infertility that is not rescued by the WW-binding protein PAWP. Hum. Mol. Genet. 25, 878-891. · DOI
  10. Ferrer-Vaquer, A., Barragan, M., Freour, T., Vernaeve, V. and Vassena, R. (2016). PLCzeta sequence, protein levels, and distribution in human sperm do not correlate with semen characteristics and fertilization rates after ICSI. J. Assist. Reprod. Genet. 33, 747-756. · DOI
  11. Heytens, E., Parrington, J., Coward, K., Young, C., Lambrecht, S., Yoon, S.-Y., Fissore, R. A., Hamer, R., Deane, C. M., Ruas, M. et al. (2009). Reduced amounts and abnormal forms of phospholipase C zeta (PLCzeta) in spermatozoa from infertile men. Hum. Reprod. 24, 2417-2428. · DOI
  12. Igusa, Y. and Miyazaki, S. (1986). Periodic increase of cytoplasmic free calcium in fertilized hamster eggs measured with calcium-sensitive electrodes. J. Physiol. 377, 193-205. · DOI
  13. Ito, J., Parrington, J. and Fissore, R. A. (2011). PLCzeta and its role as a trigger of development in vertebrates. Mol. Reprod. Dev. 78, 846-853. · DOI
  14. Kashir, J., Jones, C., Lee, H. C., Rietdorf, K., Nikiforaki, D., Durrans, C., Ruas, M., Tee, S. T., Heindryckx, B., Galione, A. et al. (2011). Loss of activity mutations in phospholipase C zeta (PLCzeta) abolishes calcium oscillatory ability of human recombinant protein in mouse oocytes. Hum. Reprod. 26, 3372-3387. · DOI
  15. Kashir, J., Konstantinidis, M., Jones, C., Lemmon, B., Lee, H. C., Hamer, R., Heindryckx, B., Deane, C. M., De Sutter, P., Fissore, R. A. et al. (2012). A maternally inherited autosomal point mutation in human phospholipase C zeta (PLCzeta) leads to male infertility. Hum. Reprod. 27, 222-231. · DOI
  16. Kashir, J., Buntwal, L., Nomikos, M., Calver, B. L., Stamatiadis, P., Ashley, P., Vassilakopoulou, V., Sanders, D., Knaggs, P., Livaniou, E. et al. (2017). Antigen unmasking enhances visualization efficacy of the oocyte activation factor, phospholipase C zeta, in mammalian sperm. Mol. Hum. Reprod. 23, 54-67. · DOI
  17. Kline, D. and Kline, J. T. (1992). Repetitive calcium transients and the role of calcium in exocytosis and cell cycle activation in the mouse egg. Dev. Biol. 149, 80-89. · DOI
  18. Knott, J. G., Kurokawa, M., Fissore, R. A., Schultz, R. M. and Williams, C. J. (2005). Transgenic RNA interference reveals role for mouse sperm phospholipase Czeta in triggering Ca2+ oscillations during fertilization. Biol. Reprod. 72, 992-996. · DOI
  19. Knott, J. G., Gardner, A. J., Madgwick, S., Jones, K. T., Williams, C. J. and Schultz, R. M. (2006). Calmodulin-dependent protein kinase II triggers mouse egg activation and embryo development in the absence of Ca2+ oscillations. Dev. Biol. 296, 388-395. · DOI
  20. Kurokawa, M., Sato, K., Wu, H., He, C., Malcuit, C., Black, S. J., Fukami, K. and Fissore, R. A. (2005). Functional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm. Dev. Biol. 285, 376-392. · DOI
  21. Lee, H. C., Yoon, S.-Y., Lykke-Hartmann, K., Fissore, R. A. and Carvacho, I. (2016). TRPV3 channels mediate Ca(2)(+) influx induced by 2-APB in mouse eggs. Cell Calcium 59, 21-31. · DOI
  22. Li, L., Lu, X. and Dean, J. (2013). The maternal to zygotic transition in mammals. Mol. Aspects Med. 34, 919-938. · DOI
  23. Miao, Y.-L., Stein, P., Jefferson, W. N., Padilla-Banks, E. and Williams, C. J. (2012). Calcium influx-mediated signaling is required for complete mouse egg activation. Proc. Natl. Acad. Sci. USA 109, 4169-4174. · DOI
  24. Morgan, D. J., Weisenhaus, M., Shum, S., Su, T., Zheng, R., Zhang, C., Shokat, K. M., Hille, B., Babcock, D. F. and McKnight, G. S., (2008). Tissue-specific PKA inhibition using a chemical genetic approach and its application to studies on sperm capacitation. Proc. Natl. Acad. Sci. USA 105, 20740-20745. · DOI
  25. Navarrete, F. A., Garcıá-Vaźquez, F. A., Alvau, A., Escoffier, J., Krapf, D., Sańchez-Caŕdenas, C., Salicioni, A. M., Darszon, A. and Visconti, P. E. (2015). Biphasic role of calcium in mouse sperm capacitation signaling pathways. J. Cell. Physiol. 230, 1758-1769. · DOI
  26. Nomikos, M., Kashir, J., Swann, K. and Lai, F. A. (2013). Sperm PLCzeta: from structure to Ca2+ oscillations, egg activation and therapeutic potential. FEBS Lett. 587, 3609-3616. · DOI
  27. Ozil, J.-P., Markoulaki, S., Toth, S., Matson, S., Banrezes, B., Knott, J. G., Schultz, R. M., Huneau, D. and Ducibella, T. (2005). Egg activation events are regulated by the duration of a sustained [Ca2+]cyt signal in the mouse. Dev. Biol. 282, 39-54. · DOI
  28. Parrington, J., Davis, L. C., Galione, A. and Wessel, G. (2007). Flipping the switch: how a sperm activates the egg at fertilization. Dev. Dyn. 236, 2027-2038. · DOI
  29. Phillips, K. P., Petrunewich, M. A. F., Collins, J. L., Booth, R. A., Liu, X. J. and Baltz, J. M. (2002). Inhibition of MEK or cdc2 kinase parthenogenetically activates mouse eggs and yields the same phenotypes as Mos(-/-) parthenogenotes. Dev. Biol. 247, 210-223. · DOI
  30. Ran, F. A., Hsu, P. D., Lin, C.-Y., Gootenberg, J. S., Konermann, S., Trevino, A. E., Scott, D. A., Inoue, A., Matoba, S., Zhang, Y. et al. (2013). Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell 154, 1380-1389. · DOI · Paper page
  31. Sanders, J. R. and Swann, K. (2016). Molecular triggers of egg activation at fertilization in mammals. Reproduction 152, R41-R50. · DOI
  32. Sanusi, R., Yu, Y., Nomikos, M., Lai, F. A. and Swann, K. (2015). Rescue of failed oocyte activation after ICSI in a mouse model of male factor infertility by recombinant phospholipase Czeta. Mol. Hum. Reprod. 21, 783-791. · DOI
  33. Satouh, Y., Nozawa, K. and Ikawa, M. (2015). Sperm postacrosomal WW domain- binding protein is not required for mouse egg activation. Biol. Reprod. 93, 94. · DOI
  34. Saunders, C. M., Larman, M. G., Parrington, J., Cox, L. J., Royse, J., Blayney, L. M., Swann, K. and Lai, F. A. (2002). PLC zeta: a sperm-specific trigger of Ca(2+) oscillations in eggs and embryo development. Development 129, 3533-3544.
  35. Sfontouris, I. A., Nastri, C. O., Lima, M. L. S., Tahmasbpourmarzouni, E., Raine- Fenning, N. and Martins, W. P. (2015). Artificial oocyte activation to improve reproductive outcomes in women with previous fertilization failure: a systematic review and meta-analysis of RCTs. Hum. Reprod. 30, 1831-1841. · DOI
  36. Shen, B., Zhang, W., Zhang, J., Zhou, J., Wang, J., Chen, L., Wang, L., Hodgkins, A., Iyer, V., Huang, X. et al. (2014). Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects. Nat. Methods 11, 399-402. · DOI
  37. Sobinoff, A. P., Sutherland, J. M. and McLaughlin, E. A. (2013). Intracellular signalling during female gametogenesis. Mol. Hum. Reprod. 19, 265-278. · DOI
  38. Suzuki, T., Yoshida, N., Suzuki, E., Okuda, E. and Perry, A. C. F. (2010). Full-term mouse development by abolishing Zn2+-dependent metaphase II arrest without Ca2+ release. Development 137, 2659-2669. · DOI
  39. Swann, K. and Lai, F. A. (2016). The sperm phospholipase C-zeta and Ca2+ signalling at fertilization in mammals. Biochem. Soc. Trans. 44, 267-272. · DOI
  40. Vanden Meerschaut, F., Nikiforaki, D., Heindryckx, B. and De Sutter, P., (2014). Assisted oocyte activation following ICSI fertilization failure. Reprod. Biomed. Online 28, 560-571. · DOI
  41. Wakai, T., Vanderheyden, V. and Fissore, R. A. (2011). Ca2+ signaling during mammalian fertilization: requirements, players, and adaptations. Cold Spring Harb. Perspect. Biol. 3, a006767. · DOI
  42. Whittingham, D. G. and Siracusa, G. (1978). The involvement of calcium in the activation of mammalian oocytes. Exp. Cell Res. 113, 311-317. · DOI
  43. Xu, Z., Abbott, A., Kopf, G. S., Schultz, R. M. and Ducibella, T. (1997). Spontaneous activation of ovulated mouse eggs: time-dependent effects on M- phase exit, cortical granule exocytosis, maternal messenger ribonucleic acid recruitment, and inositol 1,4,5-trisphosphate sensitivity. Biol. Reprod. 57, 743-750. · DOI
  44. Yanagimachi, R. and Chang, M. C. (1961). Fertilizable life of golden hamster ova and their morphological changes at the time of losing fertilizability. J. Exp. Zool. 148, 185-203. · DOI
  45. Yoon, S.-Y. and Fissore, R. A. (2007). Release of phospholipase C zeta and [Ca2+]i oscillation-inducing activity during mammalian fertilization. Reproduction 134, 695-704. · DOI
  46. Yoshida, N. and Perry, A. C. F. (2007). Piezo-actuated mouse intracytoplasmic sperm injection (ICSI). Nat. Protoc. 2, 296-304. · DOI

Cited by

Sorted by citations

Authors
Hachem, A., Godwin, J., Ruas, M., Lee, H. C., Ferrer Buitrago, M., Ardestani, G., Bassett, A., Fox, S., Navarrete, F., de Sutter, P., Heindryckx, B., Fissore, R., & Parrington, J.
Affiliations
  • University of Oxford (GB)
  • University of Massachusetts Amherst (US)
  • Ghent University Hospital 4 Department for Reproductive Medicine , , De Pintelaan 185, 9000 Ghent , Belgium
  • Ghent University Hospital (BE)
Corresponding author
Parrington, John
ORCID
Venue
Development
Publisher
The Company of Biologists
Published
2017-08-15
Volume
144
Issue
16
Pages
2914-2924
ISSN
  • 0950-1991 (ISSN-L)
  • 1477-9129
DOI
10.1242/dev.150227
PMC
PMC5592814
OpenAlex
W2724700374
Cited by
157
References
0
Citations by year
2026: 9 · 2025: 13 · 2024: 22 · 2023: 18 · 2022: 14 · 2021: 11 · 2020: 28 · 2019: 13 · 2018: 22 · 2017: 6 · 2012: 1
FWCI
17.72
Type
journal article
Language
en
Open access
Licence
www.biologists.com/user-licence-1-1/ (vor, 184 d)
Locations
  • Development (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Ghent University Academic Bibliography (Ghent University) (publishedVersion)
  • Ghent University Academic Bibliography (Ghent University) (submittedVersion, other-oa)
  • HighWire Press Open Archive (submittedVersion)
Funders
  • Iraqi Higher Committee for Education Development
  • Society for Reproduction and Fertility
  • Fonds Wetenschappelijk Onderzoek (G060615N)
  • National Institutes of Health (HD051872)
Topics
  • Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Reproductive Biology and Fertility
  • Health Sciences › Medicine › Reproductive Medicine › Sperm and Testicular Function
  • Health Sciences › Medicine › Reproductive Medicine › Reproductive Health and Technologies
Keywords
PLC, PLCZ1, calcium oscillations, oocyte activation, fertilisation (biology), polyspermy, male infertility, embryonic development
MeSH terms
Gene Editing, Animals, Calcium, Male, Mammals, Mice, Mutant Strains, Spermatogenesis, Embryonic Development, Mice, Phosphoinositide Phospholipase C, CRISPR-Cas Systems

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