Abstract：
The complex interactome crucial for successful pregnancy is constituted by the intricate network of endocrine and paracrine signaling pathways, involving gametes, embryos, and the female reproductive tract. Specifically, the oviduct exhibits distinct responses to gametes and early embryos during particular phases of the estrus cycle, a process tightly regulated by reproductive hormones. Moreover, these hormones play a pivotal role in orchestrating cyclical changes within oviductal epithelial cells. To unravel the molecular mechanisms underlying these dynamic changes, our study aimed to investigate the involvement of protein kinase A (PKA) in oviductal epithelial cells throughout the estrus cycle and in advanced pregnancy, extending our studies to oviductal epithelial cell in primary culture. By a combination of 2D-gel electrophoresis, Western blotting, and mass spectrometry, we identified 17 proteins exhibiting differential phosphorylation status mediated by PKA. Among these proteins, we successfully validated the phosphorylation status of heat shock 70 kDa protein (HSP70), aconitase 2 (ACO2), and lamin B1 (LMNB1). Our findings unequivocally demonstrate the dynamic regulation of PKA throughout the estrus cycle in oviductal epithelial cells. Also, analysis by bioinformatics tools suggest its pivotal role in mediating cyclical changes possibly through modulation of apoptotic pathways. This research sheds light on the intricate molecular mechanisms underlying reproductive processes, with implications for understanding fertility and reproductive health.
摘要：
对成功怀孕至关重要的复杂相互作用组是由复杂的内分泌和旁分泌信号通路网络构成的。涉及配子，胚胎,和女性生殖道。具体来说,输卵管在发情周期的特定阶段对配子和早期胚胎表现出不同的反应，由生殖激素严格调节的过程。此外，这些激素在调节输卵管上皮细胞的周期性变化中起着关键作用。为了解开这些动态变化背后的分子机制，我们的研究旨在探讨蛋白激酶A（PKA）在整个发情周期和晚期妊娠中输卵管上皮细胞的参与，将我们的研究扩展到原代培养的输卵管上皮细胞。通过二维凝胶电泳的组合，西方印迹,和质谱，我们鉴定了17种显示PKA介导的差异磷酸化状态的蛋白质.在这些蛋白质中，我们成功地验证了热休克70kDa蛋白（HSP70）的磷酸化状态，乌头酸酶2(ACO2),和内皮素B1(LMNB1)。我们的发现明确证明了PKA在整个输卵管上皮细胞发情周期中的动态调节。此外，生物信息学工具的分析表明，其在介导周期性变化中的关键作用可能通过调节凋亡途径。这项研究揭示了生殖过程背后复杂的分子机制，对理解生育和生殖健康有影响。
