饲料研究2025，Vol.48Issue(6)：71-77,7.DOI:10.13557/j.cnki.issn1002-2813.2025.06.014
蛋鸡GABRA1基因shRNA干扰载体构建及其对等级前卵泡颗粒细胞分子调控作用
Construction of shRNA interference vector for GABRA1 gene in laying hens and its molecular regulatory effects on prehierarchical follicle granulosa cells
摘要
Abstract
The study aimed to investigate the regulatory role of the γ-aminobutyric acid type A receptor subunit α1(GABRA1)gene in prehierarchical follicle granulosa cells of laying hens.Using targeted short hairpin RNA(shRNA)interference technology,an interference expression vector was constructed.The experiment was divided into three groups:Blank control(BC group),negative control(NC group),and shRNA GABRA1 interference group(shRNA GABRA1 group).The vectors were transfected into prehierarchical follicle granulosa cells(4～8 mm)of laying hens.After 48 hours of transfection,real-time quantitative PCR(RT-qPCR)and Western blot were used to analyze the mRNA and protein expression levels of genes related to cell proliferation(CCND1),cell differentiation,steroid synthesis genes,steroid synthesis rapid regulatory protein(StAR)cytochrome P450 family 11 subfamily A member 1(CYP11A1),apoptosis genes caspase-3(Caspase-3),and B-cell lymphoma/leukemia-2(Bcl-2).Additionally,EDU assay and flow cytometry were employed to assess cell proliferation and apoptosis.The results showed that,compared to the BC and NC groups,the mRNA and protein expression levels of CCND1,StAR,CYP11A1,and Bcl-2 were extremely upregulated in the shRNA GABRA1 group(P＜0.01),while the expression of Caspase-3 was extremely downregulated(P＜0.01).The proliferation index of granulosa cells significantly increased(P＜0.05),and the total apoptosis rate significantly decreased(P＜0.05).The study shows that GABRA1 can inhibit the proliferation and differentiation of early follicular granulosa cells,promote apoptosis,and then affect the growth and development of follicles.关键词
shRNA干扰/GABRA1/颗粒细胞/增殖/凋亡Key words
shRNA interference/GABRA1/granulosa cells/proliferation/apoptosis分类
农业科技引用本文复制引用
陈晓霞,杨辉,邹贤群,余静,刘永杰..蛋鸡GABRA1基因shRNA干扰载体构建及其对等级前卵泡颗粒细胞分子调控作用[J].饲料研究,2025,48(6):71-77,7.基金项目
吉林省科技发展计划项目(项目编号:YDZJ202201ZYTS609) （项目编号:YDZJ202201ZYTS609）
