牦牛ATF4基因组织表达与雌性生殖器官中定位分析  

Tissue expression and localization of ATF4 in female reproductive organs of yak

作　　者：蒋旭东 邬建飞 刘宇 胡双阁 马瑶 龚三你 卢建远[1] 字向东[1] JIANG Xu-dong;WU Jian-fei;LIU Yu;HU Shuang-ge;MA Yao;GONG San-ni;LU Jian-yuan;ZI Xiang-dong(The Key Laboratory of Animal Science of National Ethnic Affairs Commission,Southwest Minzu University,Chengdu 610041,China)

机构地区：[1]西南民族大学动物科学国家民委重点实验室,四川成都610041

出　　处：《西南民族大学学报（自然科学版）》2022年第2期125-134,共10页Journal of Southwest Minzu University(Natural Science Edition)

基　　金：国家重点研发计划项目(2018YFD0502303);西南民族大学中央高校基本科研业务专项资金项目(2021PTJS26)。

摘　　要：旨在克隆牦牛转录激活因子4(ATF4)基因,并检测其在不同组织的表达及雌性生殖器官中定位分析,探索ATF4在母牦牛繁殖的影响.以牦牛为实验对象,利用RT-PCR克隆得到牦牛ATF4基因CDS序列,生物信息学分析牦牛CDS区,RT-qPCR检测ATF4基因在牦牛不同组织中的表达模式,免疫组化(IHC)分析ATF4蛋白在雌性生殖器官中定位.结果表明,获得到牦牛ATF4基因cDNA序列,编码区全长1 047 bp,编码348个氨基酸,ATF4蛋白属于酸性亲水不稳定蛋白.RT-qPCR显示,ATF4基因在检测的8个组织中均有表达,在子宫和卵巢的表达量显著高于其它组织(P<0.05);妊娠期子宫、卵巢和输卵管中ATF4基因的表达量显著高于空怀期(P<0.05).IHC显示ATF4蛋白主要在牦牛卵巢的颗粒细胞、卵泡液、输卵管黏膜上皮、子宫内膜和子宫基质细胞中表达.这些结果说明ATF4基因在动物进化中比较保守,组织表达广泛,并在子宫和卵巢高表达,可能在牦牛繁殖调控中发挥重要作用.This paper cloned yak(Bos grunniens)transcription activating factor 4(ATF4)gene,and detecteds its expression in different organizations and positioning analysis in the female reproductive organs in order to explore ATF4 influence on female yak breeding.The CDS sequence of yak ATF4 gene was cloned by RT-PCR,and analyzed by bioinformatics,the expression of ATF4 gene in different yak tissues was detected by RT-qPCR,and immunohistochemistry(IHC)was used to assay ATF4 protein localization in the female reproductive organs.The results showed that the CDS sequence of yak ATF4 gene was obtained with coding region of 1047 bp which encoded 348 amino acids,an acid hydrophilic unstable protein.RT-PCR showed that ATF4 gene was expressed in all eight tissues examined.It expressed higher in uterus and ovary than in other tissues(P<0.05).The expression levels were higher in uterus,ovary and oviduct of pregnant yaks than those of non-pregnant yaks(P<0.05).IHC showed that ATF4 protein was mainly expressed in granulosa cells,follicular fluid,oviduct mucosal epithelium,endometrium and uterine stromal cells of yaks.It is concluded that ATF4 gene is conservative in the course of animal evolution and widely expressed in different tissues,particularly in uterus and ovary.It may play an important role in the regulation of yak reproduction.

关 键 词：牦牛 ATF4基因 基因克隆 实时定量PCR 免疫组化 

分 类 号：S823.85[农业科学—畜牧学]