Integrated Transcriptomic and Functional Analyses Identify an FSH-Regulated circARHGAP21/gga-miR-146a-5p/DEAF1 Module in Chicken Granulosa Cells

Follicle-stimulating hormone (FSH) supports granulosa cell survival during follicular development, but the RNA networks involved remain incompletely understood. To investigate these networks, we integrated whole-transcriptome profiling and functional assays in cultured granulosa cells from chicken prehierarchical follicles. Transcriptomic comparisons included control, FSH and FSH plus leptomycin B (LMB), a nuclear export inhibitor. Integrated analysis of these profiles identified a candidate circARHGAP21/gga-miR-146a-5p/DEAF1 regulatory module. FSH increased circARHGAP21 and DEAF1 expression and decreased gga-miR-146a-5p abundance, whereas LMB opposed these changes in FSH-treated cells. Reporter assays showed that gga-miR-146a-5p suppressed reporter activity through the predicted sites in circARHGAP21 and the DEAF1 3′ untranslated region. Transfection with the oe-circARHGAP21 construct reduced apoptosis and increased follicle-stimulating hormone receptor (FSHR) expression. Construct transfection also increased proliferating cell nuclear antigen (PCNA) expression and decreased caspase-3 (CASP3) expression. Cotransfection with a gga-miR-146a-5p mimic attenuated these effects, whereas DEAF1 overexpression produced changes similar to those observed after oe-circARHGAP21 construct transfection. Compared with the oe-circARHGAP21 construct group, the corresponding LMB-treated group showed lower DEAF1, FSHR and PCNA expression and higher CASP3 expression and apoptosis. Together, these findings identify an FSH-responsive circARHGAP21/gga-miR-146a-5p/DEAF1 module associated with granulosa cell survival and follicle-selection-related gene expression.