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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. [ABSTRACT FROM AUTHOR]

Background: Overweight and obese women have been shown to have an increased risk of recurrent miscarriage as well as other adverse reproductive outcomes, but it is yet unclear whether this is due to an effect on the endometrium, embryo or both. The current study employs proteomic analysis to examine for a potential endometrial defect in obese and overweight women with recurrent miscarriage., Methods: Proteomic tissue analysis of 21 endometrial samples obtained In the midluteal phase from 16 women with recurrent miscarriage (obese, n=12 and lean, n=4) and 5 fertile volunteers (Obese, n=2 and Lean, n=3). Proteins were separated using 2-D gel electrophoresis and principle component analysis was used to quantitatively compare protein expression between groups. Protein spots showing significantly altered expression were identified using mass spectrometry., Results: Obese and overweight recurrent miscarriage patients had a significantly increased endometrial expression of haptoglobin compared to their lean counterparts (p=0.01). These patients also displayed a significant increase in endometrial expression of transthyretin (p=0.04) and beta- globulin (p=0.04). Principle Component Analysis (PCA) of the studied groups also demonstrated that endometrial samples could be grouped based on differences in the BMI, suggesting that obesity is an independent factor influencing endometrial protein expression., Conclusions: These findings provide preliminary evidence for an alteration in the endometrial protein profile in overweight/obese women with recurrent miscarriage mainly in the form of increased haptoglobin, an inflammatory marker associated with obesity.