The egg production performance of chickens is affected by many factors, including genetics, nutrition and environmental conditions. These factors all play a role in egg production by affecting the development of follicles. MicroRNAs (miRNAs) are important non-coding RNAs that regulate biological processes by targeting genes or other non-coding RNAs after transcription. In the animal reproduction process, miRNA is known to affect the development and atresia of follicles by regulating apoptosis and autophagy of granulosa cells (GCs). In this study, we identified potential miRNAs in the atretic follicles of broody chickens and unatretic follicles of healthy chickens. We identified gga-miR-30a-5p in 50 differentially expressed miRNAs and found that gga-miR-30a-5p played a regulatory role in the development of chicken follicles. The function of miR-30a-5p was explored through the transfection test of miR-30a-5p inhibitor and miR-30a-5p mimics. In the study, we used qPCR, western blot and flow cytometry to detect granulosa cell apoptosis, autophagy and steroid hormone synthesis. Confocal microscopy and transmission electron microscopy are used for the observation of autophagolysosomes. The levels of estradiol (E2), progesterone (P4), malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by ELISA. The results showed that miR-30a-5p showed a negative effect on autophagy and apoptosis of granulosa cells, and also contributed in steroid hormones and reactive oxygen species (ROS) production. In addition, the results obtained from the biosynthesis and dual luciferase experiments showed that Beclin1 was the target gene of miR-30a-5p. The rescue experiment conducted further confirmed that Beclin1 belongs to the miR-30a-5p regulatory pathway. In summary, after deep miRNA sequencing on healthy and atretic follicles, the results indicated that miR-30a-5p inhibits granulosa cell death by inhibiting Beclin1.

中文翻译：

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对健康和闭锁鸡卵泡的 miRNA 测序分析表明，miR-30a-5p 通过靶向 Beclin1 抑制颗粒细胞死亡

鸡的产蛋性能受多种因素影响，包括遗传、营养和环境条件。这些因素都通过影响卵泡的发育在产卵中发挥作用。MicroRNAs (miRNAs) 是重要的非编码 RNA，在转录后通过靶向基因或其他非编码 RNA 来调节生物过程。在动物繁殖过程中，已知miRNA通过调节颗粒细胞（GCs）的凋亡和自噬来影响卵泡的发育和闭锁。在这项研究中，我们在育雏鸡的闭锁卵泡和健康鸡的闭锁卵泡中鉴定了潜在的 miRNA。我们在 50 种差异表达的 miRNA 中鉴定了 gga-miR-30a-5p，发现 gga-miR-30a-5p 在鸡卵泡发育中起调节作用。通过miR-30a-5p inhibitor和miR-30a-5p mimics的转染试验探讨了miR-30a-5p的功能。在研究中，我们使用qPCR、蛋白质印迹和流式细胞仪检测颗粒细胞凋亡、自噬和类固醇激素合成。共聚焦显微镜和透射电子显微镜用于观察自噬溶酶体。ELISA法检测雌二醇(E2)、孕酮(P4)、丙二醛(MDA)和超氧化物歧化酶(SOD)的含量。结果表明，miR-30a-5p对颗粒细胞的自噬和凋亡具有负面影响，并且还有助于类固醇激素和活性氧（ROS）的产生。此外，生物合成和双荧光素酶实验结果表明，Beclin1是miR-30a-5p的靶基因。进行的拯救实验进一步证实了Beclin1属于miR-30a-5p调控通路。总之，对健康和闭锁的卵泡进行深度 miRNA 测序后，结果表明 miR-30a-5p 通过抑制 Beclin1 来抑制颗粒细胞死亡。