Towards a More Comprehensive Picture of the MicroRNA-23a/b-3p Impact on Impaired Male Fertility

Abstract

The expression levels of various genes involved in human spermatogenesis are influenced by microRNAs (miRNAs), specifically microRNA-23a/b-3p. While certain genes are essential for spermatogenesis and male germ cell function, the regulation of their expression remains unclear. This study aimed to investigate whether microRNA-23a/b-3p targets genes involved in spermatogenesis and the impact of this targeting on the expression levels of these genes in males with impaired fertility. In-silico prediction and dual-luciferase assays were used to determine the potential connections between microRNA-23a/b-3p overexpression and reduced expression levels of 16 target genes. Reverse transcription-quantitative PCR (RT-qPCR) was conducted on 41 oligoasthenozoospermic men receiving infertility treatment and 41 age-matched normozoospermic individuals to verify the lower expression level of target genes. By employing dual-luciferase assays, microRNA-23a-3p was found to directly target eight genes, namely NOL4, SOX6, GOLGA6C, PCDHA9, G2E3, ZNF695, CEP41, and RGPD1, while microRNA-23b-3p directly targeted three genes, namely SOX6, GOLGA6C, and ZNF695. The intentional alteration of the microRNA-23a/b binding site within the 3′ untranslated regions (3′UTRs) of the eight genes resulted in the loss of responsiveness to microRNA-23a/b-3p. This confirmed that NOL4, SOX6, GOLGA6C, PCDHA9, and CEP41 are direct targets for microRNA-23a-3p, while NOL4, SOX6, and PCDHA9 are direct targets for microRNA-23b-3p. The sperm samples of oligoasthenozoospermic men had lower expression levels of target genes than age-matched normozoospermic men. Correlation analysis indicated a positive correlation between basic semen parameters and lower expression levels of target genes. The study suggests that microRNA-23a/b-3p plays a significant role in spermatogenesis by controlling the expression of target genes linked to males with impaired fertility and has an impact on basic semen parameters.

Oxidative Stress Biomarkers in Male Infertility: Established Methodologies and Future Perspectives

“…miR-23b-3p and miR-320a-3p can alter the expression of many specific genes in spermatogenesis such as 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 4 (PFKFB4), hyaluronan mediated motility receptor (HMMR), spermatogenesis associated 6 (SPATA6), testis expressed 15, meiosis and synapsis associated (TEX15), SRY-box transcription factor 6 (SOX6), and nucleolar protein 4 (NOL4). Being involved in the spermatogenesis process, their altered expression due to an increased aberrant expression of miRNA could compromise male fertility [136].…”