miR‐130a promotes immature porcine Sertoli cell growth by activating SMAD5 through the TGF‐β‐PI3K/AKT signaling pathway

Sertoli cells play vital roles in normal spermatogenesis, and microRNAs (miRNAs) participate in regulating Sertoli cell development. However, the functions and mechanisms of action of most identified miRNAs in porcine Sertoli cells remain largely unknown. Herein, we primarily explored the regulatory roles of miR‐130a in immature porcine Sertoli cells using EdU‐based high‐content screening assay. The results demonstrated that 27 miRNAs have potential roles in the promotion of immature porcine Sertoli cell proliferation, and miR‐130a was identified as a promising candidate. miR‐130a promoted cell cycle progression and cell proliferation, whereas it impeded cell apoptosis in immature porcine Sertoli cells. It also contributed to Sertoli cell proliferation and testis development in vivo. A TMT‐based proteomics approach revealed that miR‐130a regulated the expression of 91 proteins and multiple pathways, including the TGF‐β and PI3K/AKT signaling. miR‐130a did not directly target the 3′‐UTR of SMAD5; however, it increased SMAD5 phosphorylation. Moreover, miR‐130a enhanced TGF‐β signaling by activating SMAD5 protein, and TGF‐β signaling further activated the PI3K/AKT signaling pathway to promote cell proliferation and inhibit cell apoptosis in porcine immature Sertoli cells. Collectively, miR‐130a promoted immature porcine Sertoli cell growth by activating SMAD5 through the TGF‐β‐PI3K/AKT signaling pathway. This study, therefore, provides novel insights into the effects of miR‐130a on porcine spermatogenesis through the regulation of immature Sertoli cell proliferation and apoptosis.