Skip to main content
SpringerLink
Log in

Protein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

This study was conducted to determine if expression of the testis-specific phospholipase C Zeta1 (PLCZ1) correlated with low success or fertilization failure after ICSI in patients with normal parameters after standard semen analysis (SA).

Methods

Couples <43 years with one or two failed or low fertilization ICSI cycles. Standard Semen Analysis (SA) was performed to determine sperm parameters in male partners, whereas females were evaluated for antral follicle counts (AFC), day 3 FSH levels and peak Estradiol (E2) levels. The presence of PLCZ1 in sperm was ascertained using Western blotting and Immunofluorescence (IF) analysis. The ability of sperm to initiate changes in the intracellular concentrations of free calcium ([Ca2+]i), which is characteristic of mammalian sperm, was performed after injection of human sperm into mouse eggs loaded with the Ca2+ sensitive dye fura-2 AM.

Results

Male partners of couples with failed or low success ICSI fertilization but with normal SA parameters showed low expression levels of PLCZ1 as determined by western blotting and reduced fluorescent signal during IF studies. In addition, fewer of these males’ sperm showed PLCZ1 expression and were able to initiate robust [Ca2+]i oscillations upon injection into eggs.

Conclusion

Our data suggest that in patients with normal SA parameters but with repeated low fertilization or outright failed fertilization results after ICSI, abnormal PLCZ1 function should be considered as the underlying mechanism responsible for the failure of fertilization.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Jones K. Ca2+ oscillations in the activation of the egg and development of the embryo in mammals. Int J Dev Biol. 1998;42(1):1–10.

    CAS  PubMed  Google Scholar 

  2. Miyazaki S, Ito M. Calcium signals for egg activation in mammals. J Pharmacol Sci. 2006;100(5):545–52.

    Article  CAS  PubMed  Google Scholar 

  3. Berridge M, Irvine R. Inositol phosphates and cell signalling. Nature. 1989;341(6239):197–205. doi:10.1038/341197a0.

    Article  CAS  PubMed  Google Scholar 

  4. Berridge M. The endoplasmic reticulum: a multifunctional signaling organelle. Cell Calcium. 2002;32(5–6):235–49.

    Article  CAS  PubMed  Google Scholar 

  5. Ducibella T, Huneau D, Angelichio E, Xu Z, Schultz R, Kopf G, et al. Egg-to-embryo transition is driven by differential responses to Ca2+ oscillation number. Dev Biol. 2002;250(2):280–91. doi:10.1006/dbio.2002.0788.

    Article  CAS  PubMed  Google Scholar 

  6. Ducibella T, Fissore R. The roles of Ca2+ downstream protein kinases, and oscillatory signaling in regulating fertilization and the activation of development. Dev Biol. 2008;315(2):257–79. doi:10.1016/j.ydbio.2007.12.012.

    Article  CAS  PubMed  Google Scholar 

  7. Saunders C, Larman M, Parrington J, Cox L, Royse J, Blayney L, et al. PLC zeta: a sperm-specific trigger of Ca2+ oscillations in eggs and embryo development. Development. 2002;129(15):3533–44.

    CAS  PubMed  Google Scholar 

  8. Kurokawa M, Sato K-I, Wu H, He C, Malcuit C, Black S, et al. Functional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm. Dev Biol. 2005;285(2):376–92. doi:10.1016/j.ydbio.2005.06.029.

    Article  CAS  PubMed  Google Scholar 

  9. Kurokawa M, Sato K-I, Fissore R. Mammalian fertilization: from sperm factor to phospholipase Czeta. Biol Cell Auspices Eur Cell Biol Organ. 2004;96(1):37–45. doi:10.1016/j.biolcel.2003.11.003.

    CAS  Google Scholar 

  10. Rogers N, Hobson E, Pickering S, Lai F, Braude P, Swann K. Phospholipase Czeta causes Ca2+ oscillations and parthenogenetic activation of human oocytes. Reproduction. 2004;128(6):697–702. doi:10.1530/rep.1.00484.

    Article  CAS  PubMed  Google Scholar 

  11. Kurokawa M, Yoon S, Alfandari D, Fukami K, Sato K-I, Fissore R. Proteolytic processing of phospholipase Czeta and [Ca2+]i oscillations during mammalian fertilization. Dev Biol. 2007;312(1):407–18.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Sousa M, Tesarik J. Ultrastructural analysis of fertilization failure after intracytoplasmic sperm injection. Hum Reprod. 1994;9(12):2374–80.

    CAS  PubMed  Google Scholar 

  13. Flaherty S, Payne D, Matthews C. Fertilization failures and abnormal fertilization after intracytoplasmic sperm injection. Hum Reprod. 1998;13 Suppl 1:155–64.

    Article  PubMed  Google Scholar 

  14. Yoon S-Y, Jellerette T, Salicioni A, Lee H, Yoo M-S, Coward K, et al. Human sperm devoid of PLC zeta 1 fail to induce Ca2+ release and are unable to initiate the first step of embryo development. J Clin Investig. 2008;118(11):3671–81. doi:10.1172/jci36942.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Heytens E, Parrington J, Coward K, Young C, Lambrecht S, Yoon SY, et al. Reduced amounts and abnormal forms of phospholipase C zeta (PLCzeta) in spermatozoa from infertile men. Hum Reprod. 2009;24(10):2417–28. doi:10.1093/humrep/dep207.

    Article  CAS  PubMed  Google Scholar 

  16. Kashir J, Konstantinidis M, Jones C, Lemmon B, Lee H, Hamer R, et al. A maternally inherited autosomal point mutation in human phospholipase C zeta (PLCζ) leads to male infertility. Hum Reprod. 2012;27(1):222–31. doi:10.1093/humrep/der384.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Heindryckx B, Van der Elst J, De Sutter P, Dhont M. Treatment option for sperm- or oocyte-related fertilization failure: assisted oocyte activation following diagnostic heterologous ICSI. Hum Reprod. 2005;20(8):2237–41. doi:10.1093/humrep/dei029.

    Article  CAS  PubMed  Google Scholar 

  18. Arslan M, Bocca S, Mirkin S, Barroso G, Stadtmauer L, Oehninger S. Controlled ovarian hyperstimulation protocols for in vitro fertilization: two decades of experience after the birth of Elizabeth Carr. Fertil Steril. 2005;84(3):555–69. doi:10.1016/j.fertnstert.2005.02.053.

    Article  CAS  PubMed  Google Scholar 

  19. Hsu A, Arny M, Knee A, Bell C, Cook E, Novak A, et al. Antral follicle count in clinical practice: analyzing clinical relevance. Fertil Steril. 2011;95(2):474–9. doi:10.1016/j.fertnstert.2010.03.023.

    Article  PubMed  Google Scholar 

  20. Wakai T, Vanderheyden V, Yoon S-Y, Cheon B, Zhang N, Parys J, et al. Regulation of inositol 1,4,5-trisphosphate receptor function during mouse oocyte maturation. J Cell Physiol. 2012;227(2):705–17. doi:10.1002/jcp.22778.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Kimura Y, Yanagimachi R. Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring. Development. 1995;121(8):2397–405.

    CAS  PubMed  Google Scholar 

  22. Kurokawa M, Fissore R. ICSI-generated mouse zygotes exhibit altered calcium oscillations, inositol 1,4,5-trisphosphate receptor-1 down-regulation, and embryo development. Mol Hum Reprod. 2003;9(9):523–33. doi:10.1093/molehr/gag072.

    Article  CAS  PubMed  Google Scholar 

  23. Esfandiari N, Javed M, Gotlieb L, Casper R. Complete failed fertilization after intracytoplasmic sperm injection–analysis of 10 years data. Int J Fertil Women's Med. 2005;50(4):187–92.

    Google Scholar 

  24. Kashir J, Heindryckx B, Jones C, De Sutter P, Parrington J, Coward K. Oocyte activation, phospholipase C zeta and human infertility. Hum Reprod Updat. 2010;16(6):690–703. doi:10.1093/humupd/dmq018.

    Article  CAS  Google Scholar 

  25. Terada Y, Hasegawa H, Takahashi A, Ugajin T, Yaegashi N, Okamura K. Successful pregnancy after oocyte activation by a calcium ionophore for a patient with recurrent intracytoplasmic sperm injection failure, with an assessment of oocyte activation and sperm centrosomal function using bovine eggs. Fertil Steril. 2009;91(3). doi:10.1016/j.fertnstert.2008.09.043.

  26. Taylor S, Yoon S, Morshedi M, Lacey D, Jellerette T, Fissore R, et al. Complete globozoospermia associated with PLCζ deficiency treated with calcium ionophore and ICSI results in pregnancy. Reprod Biomed Online. 2010;20(4):559–64. doi:10.1016/j.rbmo.2009.12.024.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Escalier D. Failure of differentiation of the nuclear-perinuclear skeletal complex in the round-headed human spermatozoa. Int J Dev Biol. 1990;34(2):287–97.

    CAS  PubMed  Google Scholar 

  28. Coutton C, Zouari R, Abada F, Ben Khelifa M, Merdassi G, Triki C, et al. MLPA and sequence analysis of DPY19L2 reveals point mutations causing globozoospermia. Hum Reprod. 2012;27(8):2549–58. doi:10.1093/humrep/des160.

    Article  CAS  PubMed  Google Scholar 

  29. Sutovsky P, Oko R, Hewitson L, Schatten G. The removal of the sperm perinuclear theca and its association with the bovine oocyte surface during fertilization. Dev Biol. 1997;188(1):75–84. doi:10.1006/dbio.1997.8618.

    Article  CAS  PubMed  Google Scholar 

  30. Pierre V, Martinez G, Coutton C, Delaroche J, Yassine S, Novella C, et al. Absence of Dpy19l2, a new inner nuclear membrane protein, causes globozoospermia in mice by preventing the anchoring of the acrosome to the nucleus. Development. 2012;139(16):2955–65. doi:10.1242/dev.077982.

    Article  CAS  PubMed  Google Scholar 

  31. Kashir J, Konstantinidis M, Jones C, Heindryckx B, De Sutter P, Parrington J, et al. Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells. Fertil Steril. 2012;98(2):423–31. doi:10.1016/j.fertnstert.2012.05.002.

    Article  CAS  PubMed  Google Scholar 

  32. Kashir J, Jones C, Mounce G, Ramadan W, Lemmon B, Heindryckx B, et al. Variance in total levels of phospholipase C zeta (PLC-ζ) in human sperm may limit the applicability of quantitative immunofluorescent analysis as a diagnostic indicator of oocyte activation capability. Fertil Steril. 2013;99(1):107–17. doi:10.1016/j.fertnstert.2012.09.001.

    Article  CAS  PubMed  Google Scholar 

  33. Setti A, Paes de Almeida Ferreira Braga D, Iaconelli A, Aoki T, Borges E. Twelve years of MSOME and IMSI: a review. Reprod Biomed Online. 2013;27(4):338–52. doi:10.1016/j.rbmo.2013.06.011.

    Article  PubMed  Google Scholar 

  34. Kashir J, Sermondade N, Sifer C, Oo S, Jones C, Mounce G, et al. Motile sperm organelle morphology evaluation-selected globozoospermic human sperm with an acrosomal bud exhibits novel patterns and higher levels of phospholipase C zeta. Hum Reprod. 2012;27(11):3150–60. doi:10.1093/humrep/des312.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Baystate Medical Center, Western Campus of Tufts University School of Medicine, Springfield, MA, USA

    Margaret Arny, Daniel Grow & Teru Jellerette-Nolan

  2. Graduate Program in Animal Biotechnology and Biomedical Sciences, Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, USA

    Hoi Chang Lee & Rafael A. Fissore

  3. Department of Obstetrics and Gynecology, University of California, Los Angeles (UCLA), Los Angeles, CA, USA

    Daniel Dumesic

Authors
  1. Hoi Chang Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Margaret Arny
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Daniel Grow
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daniel Dumesic
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rafael A. Fissore
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Teru Jellerette-Nolan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Teru Jellerette-Nolan.

Additional information

Capsule Sperm with normal semen parameters from male partner of couples with repeat low or failed fertilization results after ICSI, showed low PLCZ1 expression and initiated decreased calcium release upon injection into eggs. Our data suggests PLCZ1 function could be the underlying cause and should be assessed in cycles with low or failed fertilization.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lee, H.C., Arny, M., Grow, D. et al. Protein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters. J Assist Reprod Genet 31, 749–756 (2014). https://doi.org/10.1007/s10815-014-0229-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-014-0229-9

Keywords

Search

Navigation