miRNA sequencing analysis of healthy and atretic follicles of chickens revealed that miR-30a-5p inhibits granulosa cell death via targeting Beclin1  

作　　者：Haorong He Dongmei Li Yongtong Tian Qinyao Wei Felix Kwame Amevor Congjiao Sun Chunlin Yu Chaowu Yang Huarui Du Xiaosong Jiang Menggen Ma Can Cui Zhichao Zhang Kai Tian Yao Zhang Qing Zhu Huadong Yin 

机构地区：[1]Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province,Sichuan Agricultural University,Chengdu 611130,Sichuan,China [2]National Engineering Laboratory for Animal Breeding and Key Laboratory of Animal Genetics,Breeding and Reproduction,Ministry of Agriculture and Rural Affairs,College of Animal Science and Technology,China Agricultural University,Beijing 100193,China [3]Animal Breeding and Genetics key Laboratory of Sichuan Province,Sichuan Animal Science Academy,Chengdu 610066,China [4]College of Resources,Sichuan Agricultural University,Chengdu 611130,China

出　　处：《Journal of Animal Science and Biotechnology》2022年第5期1351-1372,共22页畜牧与生物技术杂志（英文版）

基　　金：financially supported by Sichuan Science and Technology Program(2021YFYZ0031,2021YFYZ0007);China Agriculture Research System of MOF and MARA(CARS-40).

摘　　要：Background: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).Results: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.Conclusions: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.

关 键 词：Apoptosis Autophagy BECLIN1 Chicken granulosa cells MiR-30a-5p Oxidative stress RNA-seq Steroid hormone 

分 类 号：S831[农业科学—畜牧学]