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

Document Type : Original Article

Authors

1 Department of Anatomy, Faculty of Medicine, Kurdistan University of Medical Sciences, Sannandaj, Iran

2 Department of Midwifery, Falavarjan Branch, Islamic Azad University, Isfahan, Iran

Abstract

Background: Phospholipase C zeta (PLC-ζ) deficiency in sperm can underlie oocyte activation failure after intracytoplasmic
sperm injection (ICSI). The aim of this study was to determine PLC-ζ expression and location in individual
spermatozoa in each host score so that a hypo-osmotic swelling test (HOST) may be used to help routine sperm selection
for ICSI.
Materials and Methods: In this experimental study, fresh semen samples were randomly obtained from 30 men who
were 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 detected
according to World Health Organization (WHO) criteria. Then, the PLC-ζ protein localization pattern was assessed by
quantitative Immunofluorescence in individual sperm Host grades. Moreover, the sperm content of PLC-ζ protein was
evaluated by flow cytometry correlated with semen analysis parameters.
Results: In the present study, quantitive immunofluorescence analysis indicated that sperm from different host grades
exhibited 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, and
A+EQ+PA.
The sperm from HOST grade 'd' exhibited significantly higher PLC-ζ (A+PA) and (A+EQ+PA) staining compared to sperm
from 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 confirmed
that 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 exhibiting
a higher level of PLC-ζ expression.

Keywords

Main Subjects

References

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