The coordinated proliferation and apoptosis of granulosa cells plays a critical role in follicular development. To identify the exact mechanisms of how stress-driven glucocorticoid production suppresses reproduction, granulosa cells were isolated from chicken follicles at different developmental stages and then treated with corticosterone. Using CCK-8, EDU and TUNEL assays, we showed that corticosterone could trigger both anti-proliferative and pro-apoptotic effects in granulosa cells from 6 to 8 mm follicles only, while depicting no influence on granulosa cells from any preovulatory follicles. High-throughput transcriptomic analysis identified 1362 transcripts showing differential expression profiles in granulosa cells from 6 to 8 mm follicles after corticosterone treatment. Interestingly, Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that 17 genes were enriched in the TGF-β signaling pathway, and 13 showed differential expression patterns consistent with corticosterone-induced effects. The differential expression profiles of these 13 genes were examined by quantitative real-time PCR in cultured chicken ovarian granulosa cells at diverse developmental stages following corticosterone challenge for a short (8 h) or long period (24 h). After 24 h of treatment, INHBB, FST, FMOD, NOG, ACVR1, SMAD1 and ID3 were the genes that responded consistently with corticosterone-induced proliferative and apoptotic events in all granulosa cells detected. However, only ACVR1, SMAD1 and ID3 could initiate coincident expression patterns after being treated for 8 h, suggesting their significance in corticosterone-mediated actions. Collectively, these findings indicate that corticosterone can inhibit proliferation and cause apoptosis in chicken ovarian prehierarchical, but not preovulatory granulosa cells, through impeding ACVR1-SMAD1-ID3 signaling presumptively.

颗粒细胞的协同增殖和凋亡在卵泡发育中起关键作用。为了确定压力驱动的糖皮质激素产生如何抑制繁殖的确切机制，从处于不同发育阶段的鸡卵泡中分离出颗粒细胞，然后用皮质酮处理。使用 CCK-8、EDU 和 TUNEL 分析，我们发现皮质酮仅可在 6 至 8 mm 卵泡的颗粒细胞中引发抗增殖和促凋亡作用，而对任何排卵前卵泡的颗粒细胞没有影响。高通量转录组分析鉴定了 1362 个转录本，在皮质酮治疗后 6 至 8 mm 卵泡的颗粒细胞中显示出差异表达谱。有趣的是，京都基因百科全书和基因组富集分析显示，17 个基因在 TGF-β 信号通路中富集，13 个基因表现出与皮质酮诱导作用一致的差异表达模式。这 13 个基因的差异表达谱通过定量实时 PCR 在培养的鸡卵巢颗粒细胞中进行了检测，这些细胞在皮质酮攻击后的不同发育阶段进行了短期（8 小时）或长期（24 小时）的攻击。治疗24小时后，这 13 个基因的差异表达谱通过定量实时 PCR 在培养的鸡卵巢颗粒细胞中进行了检测，这些细胞在皮质酮攻击后的不同发育阶段进行了短期（8 小时）或长期（24 小时）的攻击。治疗24小时后，这 13 个基因的差异表达谱通过定量实时 PCR 在培养的鸡卵巢颗粒细胞中进行了检测，这些细胞在皮质酮攻击后的不同发育阶段进行了短期（8 小时）或长期（24 小时）的攻击。治疗24小时后，INHBB、FST、FMOD、NOG、ACVR1、SMAD1和ID3是在检测到的所有颗粒细胞中与皮质酮诱导的增殖和凋亡事件一致响应的基因。然而，只有ACVR1、SMAD1和ID3在处理 8 小时后可以启动一致的表达模式，这表明它们在皮质酮介导的作用中具有重要意义。总的来说，这些研究结果表明，皮质酮可以通过阻止 ACVR1-SMAD1-ID3 信号传导来抑制鸡卵巢分级前而非排卵前颗粒细胞的增殖并导致细胞凋亡。