绵羊生殖器官GDF9基因的表达研究  

Expression of GDF9 gene in sheep reproductive organs

作　　者：李佳蓉[1,2] 贾超 陈洋[1,2] 姜怀志 LI Jia-rong;JIA Chao;CHEN Yang;JIANG Huai-zhi(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;Key Laboratory of Animal Production,Product Quality and Security,Ministry of Education,Jilin Agricultural University,Changchun 130118,China;College of Life Engineering,Shenyang Institute of Technology,Shenyang 113122,China)

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]吉林农业大学动物生产及产品质量安全教育部重点实验室,吉林长春130118 [3]沈阳工学院生命工程学院,辽宁沈阳113122

出　　处：《中国兽医科学》2020年第8期1050-1057,共8页Chinese Veterinary Science

基　　金：吉林省科技发展计划项目(20160204018NY)。

摘　　要：生长分化因子9(GDF9)是绵羊多胎性状的重要候选基因,探讨GDF9在不同发育时期的母羊生殖器官的表达规律,对揭示其调控母羊生殖器官的发育具有重要的意义。以2月龄(性成熟前)、6月龄(性成熟)和12月龄(性成熟后)这3个不同生理阶段的雌性绵羊卵巢、输卵管、子宫内膜为对象,采用RT-PCR技术分析GDF9蛋白在相应生殖器官中的表达,并分析其相对表达量;采用免疫组织化学法对GDF9基因在相应生殖器官中的表达进行定位分析。荧光定量PCR结果显示,GDF9在2月龄与6月龄间、2月龄与12月龄间的卵巢上均表现出极显著的差异性(P<0.01),而6月龄与12月龄则无显著性差异(P>0.05);GDF9在2月龄与6月龄间的子宫内膜上表达无显著差异(P>0.05),而在2月龄与12月龄间、6月龄与12月龄间的子宫内膜上的表达均差异显著(P <0.05);GDF9在3个年龄间的输卵管上的表达均无显著性差异(P>0.05)。免疫组织化学试验结果显示,GDF9蛋白在2月龄、6月龄和12月龄绵羊子宫内膜和输卵管的上皮细胞及卵巢的颗粒细胞上均能表达,说明GDF9在不同生理阶段的绵羊生殖器官上表达且存在特异性细胞定位。Growth differentiation factor 9(GDF 9) is an important candidate gene for multiple fetal traits in sheep.To investigate the expression of GDF 9 in female sheep reproductive organs at different developmental stages is great significance to reveal its regulation on the development of female reproductive organs.In this study,the ovaries,fallopian tubes,and endometrium of female sheep at different physiological stages of 2 months(before sexual maturity),6 months(sexual maturity),and 12 months(after sexual maturity) were selected as the subjects.The expression of GDF 9 genes were analyzed by RT-PCR in the corresponding reproductive organs,and the localization of GDF9 protein was determined by immunohistochemistry in the corresponding reproductive organs.The results of RT-PCR showed that GDF 9 showed extremely significant difference in the ovaries between 2 and 6 months-old,2 and 12 months-old(P<0.01),while there was no significant difference between 6 months and 12 monthsold(P>0.05).There was no significant difference in the expression of GDF 9 in the endometrium between2 months and 6 months(P>0.05),while there was significant difference in the expression of GDF 9 in the endometrium between 2 months and 12 months,6 and 12 months-old(P<0.05). There was no significant difference in the expression of GDF 9 in fallopian tubes between the three ages(P>0.05).The results of the immunohistochemistry showed that GDF9 protein was expressed on epithelial cells of the endometrium and fallopian tubes and granulosa cells of the ovary at 2 months,6 months and 12 month.It indicates that GDF 9 is expressed on reproductive organs of sheep at different physiological stages and has specific cell localization.

关 键 词：卵巢 子宫内膜 输卵管 生长分化因子9 基因表达 定位 乾华肉用美利奴羊 

分 类 号：S857.22[农业科学—临床兽医学]