Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, 611130, China.
College of Animal Science and Technology, Southwest University, Chongqing, China.
Cell Tissue Res. 2020 Aug;381(2):337-350. doi: 10.1007/s00441-020-03207-8. Epub 2020 May 6.
Inhibin A regulates follicular development, and its expression level is related to physiological activities, such as the recruitment, selection, and predominance during follicular development. Therefore, examining inhibin A and its regulatory effects on the reproductive performance of poultry is crucial. In this study, we measured the mRNA and protein abundances of INHA and INHBA in the chicken reproductive system and determined the hormone secretion and apoptosis of follicular granulosa cells (GCs) after being treated with inhibin A protein, and flow cytometry was performed to analyze GC apoptosis in INHA-specific small RNA interference (siRNA). We detected that INHA and INHBA were mainly expressed in chicken follicles. The highest INHA mRNA abundance was found in the fifth largest preovulatory follicle (F5) (P < 0.05). INHBA mRNA expression in the largest preovulatory follicle (F1) was significantly higher than those in other follicles (P < 0.05). Similar results were found for INHA and INHBA protein expression in those follicles (P < 0.05). Treatment with inhibin A protein increased the activity of GCs in a dose-dependent manner (P < 0.05), which was characterized by decreased gene expression of pro-apoptotic factors Bax and Caspase-3 (P < 0.05) and increased expression of proliferation genes Bcl-2 and PCNA (P < 0.05). Additionally, inhibin A significantly increased the secretion of progesterone and estradiol (P < 0.05). RNAi-mediated knockdown of INHA increased apoptosis in GCs via a Caspase-3-dependent mitochondrial pathway.
抑制素 A 调节卵泡发育，其表达水平与生理活动有关，如募集、选择和优势卵泡发育。因此，研究抑制素 A 及其对家禽生殖性能的调节作用至关重要。本研究测量了鸡生殖系统中 INHA 和 INHBA 的 mRNA 和蛋白丰度，检测了抑制素 A 蛋白处理后卵泡颗粒细胞（GCs）的激素分泌和凋亡情况，并通过流式细胞术分析了 INHA 特异性小干扰 RNA（siRNA）对 GC 凋亡的影响。结果表明，INHA 和 INHBA 主要在鸡卵泡中表达。第五大卵泡（F5）中 INHA mRNA 丰度最高（P < 0.05）。最大卵泡（F1）中 INHBA mRNA 表达显著高于其他卵泡（P < 0.05）。这些卵泡中 INHA 和 INHBA 蛋白表达也有类似结果（P < 0.05）。抑制素 A 蛋白处理以剂量依赖的方式增加 GCs 的活性（P < 0.05），其特征是促凋亡因子 Bax 和 Caspase-3 的基因表达降低（P < 0.05），增殖基因 Bcl-2 和 PCNA 的表达增加（P < 0.05）。此外，抑制素 A 显著增加孕激素和雌二醇的分泌（P < 0.05）。RNAi 介导的 INHA 敲低通过 Caspase-3 依赖的线粒体途径增加 GCs 的凋亡。
