Institute of Reproductive Biology, Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.
Department of Histology and Embryology, Department of Anatomy, Poznan University of Medical Science, Poznan, Poland.
Endocrinology. 2018 Sep 1;159(9):3421-3432. doi: 10.1210/en.2018-00567.
We have previously mimicked the morphological and functional changes occurring in the oviduct epithelium during the estrous cycle in vitro by using an air-liquid interface (ALI) culture system and basolateral application of 17β-estradiol (E2) and progesterone (P4). In the current study we aimed to explore the transcriptomic changes elicited by E2 and P4 together during estrous cycle simulation and to dissect the individual effects of E2 and P4 on oviduct epithelium physiology. Primary porcine oviduct epithelial cells (POECs) (N = 6 animals) were cultured at the ALI. After differentiation for 11 days, we sequentially simulated diestrus (10 days) and estrus (2.5 days) by adding serum levels of E2 and P4 to the basolateral compartment either in combination (mix trial) or separately (P4 trial and E2 trial, respectively). Cell response was evaluated by microarray analysis (mix and P4 trials), quantitative RT-PCR, and histomorphometry (all trials). When we compared simulated diestrus with estrus stage in the mix trial, there were 169 (142 upregulated and 27 downregulated) differentially expressed genes (DEGs; fold change ≥1.5). In the P4 trial, 108 DEGs (83 upregulated and 25 downregulated) were detected. Gene enrichment analysis revealed that immune-related pathways were exclusively affected in the mix trial. In both mix and P4 trials, POECs exhibited in vivo-like morphological changes regarding epithelium height and portion of ciliated cells. However, E2 alone did not trigger morphological changes. We deduce that P4 mainly drives structural variations, and E2 is imperative for regulating immune function of the oviduct epithelium during estrous cycle.
我们之前使用气液界面（ALI）培养系统和基底外侧应用 17β-雌二醇（E2）和孕酮（P4）在体外模拟发情周期中输卵管上皮发生的形态和功能变化。在本研究中，我们旨在探索 E2 和 P4 共同在发情周期模拟过程中引起的转录组变化，并剖析 E2 和 P4 对输卵管上皮生理学的单独影响。将原代猪输卵管上皮细胞（POEC）（N = 6 只动物）在 ALI 中培养。分化 11 天后，我们通过将 E2 和 P4 的血清水平分别添加到底侧腔室（分别为 P4 试验和 E2 试验）或联合添加（混合试验）来依次模拟间情期（10 天）和发情期（2.5 天）。通过微阵列分析（混合和 P4 试验）、定量 RT-PCR 和组织形态计量学（所有试验）评估细胞反应。当我们将混合试验中的模拟间情期与发情期进行比较时，有 169 个（142 个上调和 27 个下调）差异表达基因（DEG；倍数变化≥1.5）。在 P4 试验中，检测到 108 个 DEG（83 个上调和 25 个下调）。基因富集分析表明，免疫相关途径仅在混合试验中受到影响。在混合和 P4 试验中，POECs 在关于上皮高度和纤毛细胞比例的体内样形态变化方面表现出相似性。然而，单独的 E2 并未引发形态变化。我们推断 P4 主要驱动结构变化，而 E2 对于发情周期中输卵管上皮的免疫功能调节至关重要。
