PDF(Pubmed)
Abstract：
Ovarian granulosa cells are essential to gonadotrophin-regulated estrogen production, female cycle maintenance and fertility. The epithelial Na+ channel (ENaC) is associated with female fertility; however, whether and how it plays a role in ovarian cell function(s) remained unexplored. Here, we report patch-clamp and Na+ imaging detection of ENaC expression and channel activity in both human and mouse ovarian granulosa cells, which are promoted by pituitary gonadotrophins, follicle stimulating hormone (FSH) or luteinizing hormone (LH). Cre-recombinase- and CRISPR-Cas9-based granulosa-specific knockout of ENaC α subunit (Scnn1a) in mice resulted in failed estrogen elevation at early estrus, reduced number of corpus luteum, abnormally extended estrus phase, reduced litter size and subfertility in adult female mice. Further analysis using technologies including RNA sequencing and Ca2+ imaging revealed that pharmacological inhibition, shRNA-based knockdown or the knockout of ENaC diminished spontaneous or stimulated Ca2+ oscillations, lowered the capacity of intracellular Ca2+ stores and impaired FSH/LH-stimulated transcriptome changes for estrogen production in mouse and/or human granulosa cells. Together, these results have revealed a previously undefined role of ENaC in modulating gonadotrophin signaling in granulosa cells for estrogen homeostasis and thus female fertility.
摘要：
卵巢颗粒细胞对促性腺激素调节的雌激素产生至关重要，女性周期维持和生育能力。上皮Na+通道（ENaC）与女性生育能力有关；然而，它是否以及如何在卵巢细胞功能中发挥作用仍有待探索。这里,我们报道了人和小鼠卵巢颗粒细胞中ENaC表达和通道活性的膜片钳和Na+成像检测，由垂体促性腺激素促进，卵泡刺激素（FSH）或黄体生成素（LH）。小鼠中基于Cre重组酶和CRISPR-Cas9的颗粒特异性敲除ENaCα亚基（Scnn1a）导致早期发情时雌激素升高失败，黄体数量减少，异常延长的发情期，减少成年雌性小鼠的产仔数和低生育力。使用包括RNA测序和Ca2+成像在内的技术进行的进一步分析显示，基于shRNA的敲除或ENaC的敲除减少了自发或受刺激的Ca2振荡，降低了细胞内Ca2储存的能力，并损害了FSH/LH刺激的转录组变化，从而在小鼠和/或人颗粒细胞中产生雌激素。一起,这些结果揭示了ENaC在调节颗粒细胞中的促性腺激素信号以促进雌激素稳态和女性生育能力方面的作用。
