Androgen receptor gene deficiency results in the reduction of steroidogenic potential in porcine luteal cells

Luteal steroidogenesis is critical to implantation and pregnancy maintenance in mammals. The role of androgen receptors (AR) in the progesterone (P₄) producing luteal cells of porcine corpus luteum (CL) remains unexplored. The aim of the present study was to establish <i>AR</i> gene knock out (KO) porcine luteal cell culture system model by CRISPR/Cas9 genome editing technology and to study the downstream effects of <i>AR</i> gene deficiency on steroidogenic potential and viability of luteal cells. For this purpose, genomic cleavage detection assay, microscopy, RT-qPCR, ELISA, annexin, MTT, and viability assay complemented by bioinformatics analysis were employed. There was significant downregulation (<i>p</i> &lt; 0.05) in the relative mRNA expression of steroidogenic marker genes <i>STAR, CYP11A1</i>, <i>HSD3B1</i> in <i>AR</i> KO luteal cells as compared to the control group, which was further validated by the significant (<i>p</i> &lt; 0.05) decrease in the P₄ production. Significant decrease (<i>p</i> &lt; 0.05) in relative viability on third passage were also observed. The relative mRNA expression of hypoxia related gene <i>HIF1A</i> was significantly (<i>p</i> &lt; 0.05) downregulated in <i>AR</i> KO luteal cells. Protein-protein interaction analysis mapped AR to signaling pathways associated with luteal cell functionality. These findings suggests that <i>AR</i> gene functionality is critical to luteal cell steroidogenesis in porcine.