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

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

Abstract

Purpose

Sperm-borne PLCζ protein induces Ca2+ 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.

Conclusion

Failed 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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Acknowledgments

We would like to thank the embryology laboratory technical staff from Clínica Eugin for their help in sample handling and Désirée García for statistical support.

Author information

Authors and Affiliations

  1. Clínica EUGIN, Travessera de les Corts 322, 08029, Barcelona, Spain

    Anna Ferrer-Vaquer, Montserrat Barragan, Thomas Freour, Valérie Vernaeve & Rita Vassena

  2. Service de médecine et biologie de la reproduction, CHU de Nantes, Nantes, France

    Thomas Freour

  3. INSERM UMR 1064, Nantes, France

    Thomas Freour

Authors
  1. Anna Ferrer-Vaquer
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  2. Montserrat Barragan
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  3. Thomas Freour
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  4. Valérie Vernaeve
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Corresponding author

Correspondence to Rita Vassena.

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The authors declare that they have no conflict of interest.

Additional information

Capsule PLCζ seems to be a necessary but not sufficient factor in determining the molecular pathway involved in oocyte activation.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Table 1

Sperm parameters from patients. (DOCX 15 kb)

Supplementary Table 2

Sperm parameters from donors. (DOCX 14 kb)

Supplementary Table 3

Expression of PLCz in patient samples expressed as a percentage of cells. (DOCX 26 kb)

Supplementary Table 4

Expression of PLCz in donor samples expressed as a percentage of cells. (DOCX 21 kb)

Supplementary Table 5

p values of statistical significance for comparison of distribution patters of PLCz in patients and donors. p ≤ 0.01. (DOCX 17 kb)

Supplementary Table 6

Pearson correlation between sperm parameters (motility/ concentration) and distribution patterns of PLCz within the sperm cell. (DOCX 16 kb)

Supplementary Fig. 1

Image of the full blots used to elaborate Fig. 4. Western Blots showing PLCζ and tubulin expression in total protein lysates of donor (upper panels) and patient (lower panels) samples. P = patient; D = donor. Blots were run in different days, loading D6 and D7 donors in blot from patients as an internal technical control. Red squares mark the region of the blot used to prepare Fig. 4. (GIF 331 kb)

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(TIF 1539 kb)

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Ferrer-Vaquer, A., Barragan, M., Freour, T. et al. PLCζ 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 (2016). https://doi.org/10.1007/s10815-016-0718-0

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  • DOI: https://doi.org/10.1007/s10815-016-0718-0

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