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Online First

10.4103/aja.aja_67_20

Advancing male age differentially alters levels and localization patterns of PLCzeta in sperm and testes from different mouse strains

Junaid Kashir1,2, Bhavesh V Mistry2, Maha Adel Gumssani1,2, Muhammad Rajab2, Reema Abu-Dawas1,2, Falah AlMohanna2, Michail Nomikos3, Celine Jones4, Raed Abu-Dawud2, Nadya Al-Yacoub2, Kevin Coward4, F Anthony Lai3,5, Abdullah M Assiri1,2

1 College of Medicine, Alfaisal University, Riyadh 11533, Saudi Arabia
2 Department of Comparative Medicine, King Faisal Specialist Hospital and Research Centre, Riyadh 11211, Saudi Arabia
3 College of Medicine, QU Health, Qatar University, Doha PO Box 2713, Qatar
4 Nuffield Department of Women's and Reproductive Health, University of Oxford, Level 3, Women's Centre, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom
5 Biomedical Research Centre, Qatar University, Doha PO Box 2713, Qatar

Correspondence: Dr. J Kashir (jkashir@alfaisal.edu)

13-Nov-2020

Abstract

Sperm-specific phospholipase C zeta (PLCζ) initiates intracellular calcium (Ca2+) transients which drive a series of concurrent events collectively termed oocyte activation. Numerous investigations have linked abrogation and absence/reduction of PLCζ with forms of male infertility in humans where oocyte activation fails. However, very few studies have examined potential relationships between PLCζ and advancing male age, both of which are increasingly considered to be major effectors of male fertility. Initial efforts in humans may be hindered by inherent PLCζ variability within the human population, alongside a lack of sufficient controllable repeats. Herein, utilizing immunoblotting, immunofluorescence, and quantitative reverse transcription PCR (qRT-PCR) we examined for the first time PLCζ protein levels and localization patterns in sperm, and PLCζ mRNA levels within testes, from mice at 8 weeks, 12 weeks, 24 weeks, and 36 weeks of age, from two separate strains of mice, C57BL/6 (B6; inbred) and CD1 (outbred). Collectively, advancing male age generally diminished levels and variability of PLCζ protein and mRNA in sperm and testes, respectively, when both strains were examined. Furthermore, advancing male age altered the predominant pattern of PLCζ localization in mouse sperm, with younger mice exhibiting predominantly post-acrosomal, and older mice exhibiting both post-acrosomal and acrosomal populations of PLCζ. However, the specific pattern of such decline in levels of protein and mRNA was strain-specific. Collectively, our results demonstrate a negative relationship between advancing male age and PLCζ levels and localization patterns, indicating that aging male mice from different strains may serve as useful models to investigate PLCζ in cases of male infertility and subfertility in humans.

Keywords: ageing; fertilization; male infertility; oocyte activation; phospholipase C zeta; sperm

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