HIRA基因突变对绵羊颗粒细胞中mRNA和蛋白表达及细胞激素分泌的影响  

Effects of HIRA gene mutations on mRNA,protein expression and cell hormone secretion in sheep(Ovis aries)granulosa cells

作　　者：周梅[1,2] 刘秋月 孙庆[1] 田志龙 向光明 狄冉 王翔宇[1] 储明星[1] ZHOU Mei;LIU Qiuyue;SUN Qing;TIAN Zhilong;XIANG Guangming;DI Ran;WANG Xiangyu;CHU Mingxing(Institute of Animal Science,Chinese Academy of Agricultural Sciences,Key Laboratory of Animal Genetics,Breeding and Reproduction of Ministry of Agriculture and Rural Affairs,Beijing 100193,China;Institute of Animal Husbandry and Veterinary Medicine,Anhui Academy of Agricultural Sciences,Key Laboratory of Pig Molecular Quantitative Genetics of Anhui Academy of Agricultural Sciences,Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety Engineering,Hefei 230001,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,农业农村部动物遗传育种与繁殖重点实验室,北京100193 [2]安徽省农业科学院畜牧兽医研究所,猪分子数量遗传学安徽省农业科学院重点实验室,畜禽产品安全工程安徽省重点实验室,合肥230001

出　　处：《上海农业学报》2023年第2期76-81,共6页Acta Agriculturae Shanghai

基　　金：国家转基因科技重大专项(2016ZX08009-003-006);国家肉羊产业技术体系专项(CARS-38);中国农业科学院科技创新工程(CAAS-ZDRW202106、ASTIP-IAS13);安徽省自然科学基金项目(2108085QC135)。

摘　　要：以绵羊颗粒细胞为研究对象,利用实时荧光定量PCR(quantitative real-time PCR,qPCR)和免疫荧光技术检测组蛋白细胞周期调节(histone cell cycle regulator,HIRA)基因g.71874104G>A和g.71833755T>C位点突变引起的mRNA和蛋白表达以及孕酮(progesterone,P4)和雌二醇(estradiol,E2)分泌水平在绵羊卵巢颗粒细胞中的变化。结果表明:g.71874104G>A和g.71833755T>C位点突变均导致卵巢颗粒细胞中HIRA蛋白表达发生显著变化(P<0.05),但HIRA基因的mRNA表达量均无明显差异,并且仅g.71874104G>A突变位点对P4和E2的分泌有显著影响(P<0.05)。由此推测HIRA基因的g.71874104G>A和g.71833755T>C两个突变位点通过改变绵羊卵巢颗粒细胞中HIRA蛋白表达量,进而影响后续的受精和胚胎发育过程,最终影响绵羊的生殖能力。The sheep granulosa cells were selected as the research object.Quantitative real-time PCR(qPCR)and immunofluorescence technology were performed to detect the change of mRNA and protein expression,secretion levels of progesterone(P4)and estradiol(E2)in sheep ovarian granulosa cells caused by mutations at g.71874104G>A and g.71833755T>C loci in HIRA gene.The results showed that,significant changes in the expression of HIRA protein were found in ovarian granulosa cells for both g.71874104G>A and g.71833755T>C mutations(P<0.05),while there was no obvious difference in the expression of HIRA mRNA,and only g.71874104G>A mutation had a significant effect on the secretion of P4 and E2(P<0.05).In summary,we speculated that the g.71874104G>A and g.71833755T>C mutations in HIRA gene might affect the subsequent fertilization and embryonic development mainly by changing the HIRA protein expression in sheep ovarian granulosa cells,and ultimately affect the reproduction ability of sheep.

关 键 词：绵羊 HIRA基因 蛋白表达 激素分泌 

分 类 号：S826.2[农业科学—畜牧学]