International Journal of Fertility and Sterility (Jan 2024)

Assessment of Expression Levels and Localization Patterns of Phospholipase C zeta in Different Grades of HOST in Human Sperm

  • Azra Allahveisi,
  • Elham Yousefian

DOI
https://doi.org/10.22074/ijfs.2023.1973614.1396
Journal volume & issue
Vol. 18, no. 1
pp. 26 – 31

Abstract

Read online

Background: Phospholipase C zeta (PLC-ζ) deficiency in sperm can underlie oocyte activation failure after intracytoplasmicsperm injection (ICSI). The aim of this study was to determine PLC-ζ expression and location in individualspermatozoa in each host score so that a hypo-osmotic swelling test (HOST) may be used to help routine sperm selectionfor ICSI.Materials and Methods: In this experimental study, fresh semen samples were randomly obtained from 30 men whowere referred to the Andrology Unit of the Infertility Center. Samples were processed by density gradient centrifugation(DGC) and exposed to hypotonic conditions. Seven different tail patterns, classified from ‘a’ to ‘g’ can be detectedaccording to World Health Organization (WHO) criteria. Then, the PLC-ζ protein localization pattern was assessed byquantitative Immunofluorescence in individual sperm Host grades. Moreover, the sperm content of PLC-ζ protein wasevaluated by flow cytometry correlated with semen analysis parameters.Results: In the present study, quantitive immunofluorescence analysis indicated that sperm from different host gradesexhibited seven localization patterns of PLC-ζ of acrosomal (A); equatorial (EQ), and postacrosomal (PA) patterns.A+EQ=acrosomal and equatorial, A+PA=acrosomal and post-acrosomal, EQ+PA=equatorial and post-crosomal, andA+EQ+PA.The sperm from HOST grade 'd' exhibited significantly higher PLC-ζ (A+PA) and (A+EQ+PA) staining compared to spermfrom other grades (P=0.006). The sperm from grade 'd' exhibited higher PLC-ζ (EQ+PA) compared with other grades(P=0.001). However, grade 'd' was not significantly different from 'c' (P=0.087). Analysis of the combined results confirmedthat there was a clear reduction in PLC-ζ immunofluorescence in Host grades 'a', 'f' and 'g' sperms.Conclusion: Our data suggest that HOST may represent a useful diagnostic tool for the selection of sperms exhibitinga higher level of PLC-ζ expression.

Keywords