Testosterone-Dependent Porcine miR-26a-5p and let-7g-5p Act as Signaling Mediators to Regulate Sperm Apoptosis via Targeting PTEN and PMAIP1

We found that differentially expressed (DE) miRNAs were mainly enriched in pathways involved in cell apoptosis, including mTOR and p53 signaling pathways. Furthermore, gain- and loss-of-function analyses and dual luciferase assays demonstrated that endogenous miR-26a-5p and let-7g-5p have potential anti-apoptotic and pro-survival functions in sperm cells through targeting PTEN and PMAIP1 genes. Further comparisons revealed high similarities in miRNA profiles between seminal plasma exosomes and sperm cells, both in HC and CT groups. Our study revealed that miRNAs in sperm cells and seminal plasma exosomes have important functions in male reproduction, suggesting they could be used as potential treatment targets or novel diagnostic markers for male infertility. We used small RNA sequencing to detect miRNA transcriptomes of sperm from pigs with prepubertal hemicastration (HC) and compared them with control libraries (CT) (3 vs. 3). The relationship between sperm cells and corresponding seminal plasma exosomes with respect to miRNAome was also detected.