Variance in total levels of phospholipase C zeta PLCζ in human sperm may limit the applicability of quantitative immunofluorescent analysis as a diagnostic indicator of oocyte activation capability

Capsule:
Examination of total levels and localisation patterns of PLCζ in control and patient sperm. Sperm from control and OAD males exhibited significant variance in total levels and localization patterns of PLCζ protein.

Authors:
Junaid Kashir, Ph.D., Celine Jones, Ginny Mounce, M.Sc., Walaa M. Ramadan, M.Sc., Bernadette Lemmon, B.Sc., Bjorn Heindryckx, Ph.D., Petra De Sutter, M.D., Ph.D., John Parrington, Ph.D., Karen Turner, Ph.D., Tim Child, M.D., Enda McVeigh, M.D., Kevin Coward, Ph.D.

Volume 99, Issue 1, Pages 107-117.e3, January 2013

Abstract:
Objective:
To examine whether similar levels of PLCζ protein are present in sperm from males whose ejaculates resulted in normal oocyte activation, and to examine whether a predominant pattern of PLCζ localisation is linked to normal oocyte activation ability.

Design:
Laboratory study.

Setting:
University laboratory.

Patient(s):
Control subjects (men with proven oocyte activation capacity; n = 16) and men whose sperm resulted in recurrent intracytoplasmic sperm injection failure (oocyte activation deficient [OAD]; n = 5).

Intervention(s):
Quantitative immunofluorescent analysis of PLC-ζ protein in human sperm.

Main Outcome Measure(s):
Total levels of PLC-ζ fluorescence, proportions of sperm exhibiting PLC-ζ immunoreactivity, and proportions of PLC-ζ localization patterns in sperm from control and OAD men.

Results:
Sperm from controls presented a significantly higher proportion of sperm exhibiting PLCζ immunofluorescence as opposed to infertile men diagnosed with OAD (82.6% and 27.4% respectively). Total levels of PLCζ in sperm from individual controls and OAD patients exhibited significant variance (p ≤0.05), with sperm from 10 out of 16 (62.5%) exhibiting levels comparable to OAD samples. Predominant PLCζ localisation patterns varied between control and OAD samples, with no predictable or consistent motif.

Conclusions:
The results indicate that sperm from control men exhibited significant variance in total levels of PLC-ζ protein, as well as significant variance in the predominant localization pattern. Such variance may hinder the diagnostic application of quantitative PLC-ζ immunofluorescent analysis.

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