In Vitro Mimicking of Estrous Cycle Stages in Porcine Oviduct Epithelium Cells: Estradiol and Progesterone Regulate Differentiation, Gene Expression, and Cellular Function
In Vitro Mimicking of Estrous Cycle Stages in Porcine Oviduct Epithelium Cells: Estradiol and Progesterone Regulate Differentiation, Gene Expression, and Cellular Function
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DOI：
10.1095/biolreprod.113.108829
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发表时间：
2013-09-01
影响因子：
3.6
通讯作者：
Schoen, Jennifer
中科院分区：
文献类型：
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作者：
Chen, Shuai;Einspanier, Ralf;Schoen, Jennifer
在整个动情周期中，输卵管上皮经历显着的形态和功能变化。为了阐明 17β 雌二醇 (E2) 和孕酮 (P4) 的周期性细胞事件和相关调节机制，我们使用原代猪输卵管上皮细胞 (POEC) 培养系统在体外模拟动情周期阶段。细胞在空气/液体界面中极化，然后用E2和P4处理生理时间段：在实验1中，高浓度P4和低浓度E2持续10天类似于动情间期；在实验2中，在前一次发情间期之后，连续2.5天的高E2和低P4代表发情期。组织形态计量学和电子显微镜显示在激素刺激的影响下细胞高度、细胞群和纤毛密度发生周期性变化。跨上皮电阻在模拟发情间期较高，但在发情期降低。因此，E2 和 P4 影响细胞极性、纤毛细胞和分泌细胞的转化以及输卵管上皮的电导率。模拟发情间期导致激素受体（PGR 和 ESR1）和其他上皮标记物（MUC16、OVGP1 和 HSP90B1）表达显着下降，而连续模拟发情期则导致这些标记物增加。一些标记基因的激素调节显然具有时间依赖性。此外，POEC 在模拟发情中显示出精子结合能力增加。在这项研究中，我们还提出了一种基于 AndroVision 软件的新方法，该方法可以常规用作纤毛活动的参数，并且我们首次在体外显示了沿上皮衬里的流体运动模式。
Throughout the estrous cycle the oviduct epithelium undergoes dramatic morphological and functional changes. To elucidate cyclic cellular events and associated regulation mechanisms of 17beta estradiol (E2) and progesterone (P4), we mimicked estrous cycle stages in vitro using a culture system of primary porcine oviduct epithelium cells (POEC). Cells were polarized in an air/liquid interface and then treated with E2 and P4 for physiological time periods: In experiment 1, high concentration of P4 with low concentration of E2 for 10 days resembled diestrus; in experiment 2, following the previous diestrus, sequential high E2 with low P4 for 2.5 days represented estrus. Histomorphometry and electron microscopy showed cyclic changes in cellular height, cell population, and cilia density under the influence of hormone stimulation. Transepithelial electrical resistance was high in simulated diestrus but reduced in estrus. Thus, E2 and P4 affect cellular polarity, transformation of ciliated and secretory cells, as well as electrical conductivity of oviduct epithelium. Simulation of diestrus led to significant decrease in expression of hormone receptors (PGR and ESR1) and other epithelial markers (MUC16, OVGP1, and HSP90B1), while sequential simulated estrus caused an increase in these markers. The hormonal regulation of some marker genes was clearly time-dependent. Furthermore, POEC showed increased sperm-binding capacity in simulated estrus. In this study, we also present a novel approach based on the AndroVision software, which can be routinely utilized as a parameter for ciliary activity, and for the first time, we showed fluid movement patterns along the epithelium lining in vitro.