shRNA mediated ablation of prostate and testis expressed (Pate) messenger RNA results in impaired sperm function and fertility

Abstract

Spermatogenesis and sperm maturation are complex processes mediated by a variety of proteins present in the testicular and epididymal mileu. In the recent years, functional characterization of these proteins is being studied by genetic manipulations that involve targeted over expression or knock down of specific genes. In this study, we adopted FuGENE 6-based in vivo transfection of rat cauda epididymis with pGFP-V-RS plasmid that encodes shRNA to knock down Pate mRNA levels to implicate a possible role for this gene in sperm function. The mRNA levels of Pate gene were significantly decreased in HEK cells as well as in cauda of rats upon transfection with pGFP-V-RS plasmid that encodes shRNA targeting Pate gene. Spermatozoa obtained from the transfected cauda displayed impairment in capacitation and acrosome reaction. Furthermore, rats subjected to in vivo transfection to knock down Pate mRNA levels had compromised fertility. Results of our study indicate a role for Pate gene in sperm function and can be exploited as a potential male contraceptive target.