Unveiling the role of protein kinase A (PKA) activity in bovine oviductal epithelial cells: implications on apoptotic signaling pathways during the estrous cycle

Abstract

The complex interactome crucial for successfulpregnancy is constituted by the intricate network of endocrine and paracrinesignaling pathways, involving gametes, embryos, and the female reproductivetract. Specifically, the oviduct exhibits distinct responses to gametes andearly embryos during particular phases of the estrus cycle, a process tightlyregulated by reproductive hormones. Moreover, these hormones play a pivotalrole in orchestrating cyclical changes within oviductal epithelial cells. Tounravel the molecular mechanisms underlying these dynamic changes, our studyaimed to investigate the involvement of Protein Kinase A (PKA) in oviductalepithelial cells throughout the estrus cycle and in advanced pregnancy, extendingour studies to oviductal epithelial cell in primary culture. By a combinationof 2D-gel electrophoresis, Western blotting, and mass spectrometry, weidentified 17 proteins exhibiting differential phosphorylation status mediatedby PKA. Among these proteins, we successfully validated the phosphorylationstatus of Heat shock 70 kDa protein (HSP70), Aconitase 2 (ACO2), and Lamin B1 (LMNB1).Our findings unequivocally demonstrate the dynamic regulation of PKA throughoutthe estrus cycle in oviductal epithelial cells. Also, analysis by bioinformatictools suggest its pivotal role in mediating cyclical changes possibly throughmodulation of apoptotic pathways. This research sheds light on the intricatemolecular mechanisms underlying reproductive processes, with implications forunderstanding fertility and reproductive health.