Abstract
BackgroundTo examine human microRNA expression in fertile men and subsequently to compare expression patterns of miRNAs in fertile and infertile men, specifically men with Sertoli Cell Only (SCO) histopathology.MethodsTesticular tissues from men with azoospermia and SCO, as well as those of men with normal spermatogenesis, were analyzed. MicroRNA was isolated using the miRCURY™ RNA Purification Kit. A miRCURY LNA™ Universal RT system was used for detection of microRNA by quantitative real-time PCR. MicroRNA localization was performed by in situ hybridizations (ISH) on formalin-fixed paraffin embedded (FFPE) tissue utilizing miRCURY LNA™ microRNA ISH technology. Statistical analysis was performed by GenEx V5.0.ResultsMicroRNA expression was determined for 13 normal fertile men and 5 men with the confirmed diagnosis of diffuse SCO. MiR-202-5p expression was reduced by 17-fold (P < 0.00001) in tissue from SCO men compared to normal. MiR-34c-5p was reduced by 346-fold (P < 0.00001), miR-10b was reduced 18-fold (P < 0.00001), miR-191 was reduced 20-fold (P = 0.001) and miR-126 was reduced 40-fold (P < 0.00001)) in tissues from SCO compared to normal fertile men. Using ISH, miR-202-5p was localized to Sertoli cells of men with normal spermatogenesis, but not in the Sertoli cells of men with SCO.ConclusionNumber of miRNAs are differentially expressed in normal fertile men compared to men with SCO. MicroRNA-202-5p is localized to Sertoli cells and its expression dramatically differs between fertile men and men whose germ cells are depleted, suggesting a novel interaction for regulating microRNA expression between the somatic and germ cell components of the seminiferous epithelium.
Highlights
To examine human microRNA expression in fertile men and subsequently to compare expression patterns of miRNAs in fertile and infertile men, men with Sertoli Cell Only (SCO) histopathology
In this study we quantitatively examined miRNAs expression in normal human testis and in men with a severe form of infertility: azoospermia associated with Sertoli Cell Only (SCO) syndrome
Expression of miRNA was determined for 13 normal fertile men and 5 men with the confirmed diagnosis of diffuse SCO using the 96 well plates of previously selected primers that are relevant for testicular tissue
Summary
To examine human microRNA expression in fertile men and subsequently to compare expression patterns of miRNAs in fertile and infertile men, men with Sertoli Cell Only (SCO) histopathology. Infertility affects 10%–15% of couples worldwide (WHO, 1983) [1]. Half of all infertility cases are due to male factors, and about 60–75% of male infertility is idiopathic. Most “idiopathic” male infertility is thought to be caused by yet-to-be-identified genetic defects [2]. Spermatogenesis is a multistep complex process that displays strictly a regulated spatiotemporal gene expression, and during certain aspects of germ cell division, mRNA translation is significantly repressed [3]. Studies have indicated that microRNAs (miRNAs) may play a role in translational repression during spermatogenesis [4]. The deregulation of miRNAs could play an essential role in spermatogenic dysfunction
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