GCNF在牦牛睾丸不同发育阶段中的表达与定位  

Expression and localization of GCNF in different developmental stages of yak testis

作　　者：闫翠霞 王强龙 张同享 何翃闳 潘阳阳 余四九 徐庚全 秦玉洁 YAN Cui-xia;WANG Qiang-long;ZHANG Tong-xiang;HE Hong-hong;PAN Yang-yang;YU Si-jiu;XU Geng-quan;QIN Yu-jie(Gansu Bovine,Ovine and Caprin Embryo Engineering Technology Research Center,College of Animal Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院甘肃省牛羊胚胎工程技术研究中心,甘肃兰州730070

出　　处：《中国兽医科学》2021年第3期388-394,共7页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31972760);甘肃省教育厅产业支撑引导项目(2019C-03);甘肃省教育厅创新能力提升项目(2019B-081)。

摘　　要：生殖细胞核因子(GCNF)是核受体超家族的一员。相关研究发现,GCNF在睾丸和卵巢中高表达,可能在配子减数分裂中发挥作用。为探究GCNF在牦牛睾丸中的表达与定位,采集牦牛不同发育阶段(幼年、成年、老年)的睾丸组织,采用实时荧光定量PCR(qRT-PCR)检测GCNF基因在组织中的相对表达量;采用Western-blot(WB)检测GCNF蛋白在不同年龄阶段睾丸中的表达水平;免疫组织化学方法分析GCNF在牦牛睾丸分布特征。qRT-PCR和WB检测结果显示,GCNF在牦牛睾丸不同年龄阶段均有表达,且表达存在差异性。幼年期GCNF在基因水平和蛋白水平表达量最高且显著高于成年期和老年期;老年期显著高于成年期,成年期表达量最低。免疫组织化学结果显示:在睾丸不同发育阶段GCNF的表达定位无明显差异,主要在生精细胞和圆形精子细胞中表达,间质细胞和支持细胞中也有少量表达。上述研究结果表明,GCNF可能参与精子的发生及分化变形,这为牦牛生殖生育研究提供了理论基础和新的依据。Germ cell nuclear factor(GCNF) is a member of the nuclear receptor superfamily. Related studies have found that GCNF is highly expressed in testis and ovary and may play a role in gamete meiosis. To explore the expression and localization of GCNF in yak testes,testicular tissues of Niu cattle at different developmental stages(infancy,adulthood and old age) were collected and the relative expression of GCNF gene in tissues was detected by real-time fluorescence quantitative PCR(q RT-PCR).The expression level of GCNF protein in testis of different age stages was detected by Western-blot(WB).The distribution of GCNF in yak testis was analyzed by immunized histochemical method. [results] the results of q RT-PCR and WB showed that GCNF was expressed in different age stages of bovine testis, and the expression was different.The expression of GCNF in juvenile was the highest and significantly higher than that in adulthood and senility at gene and protein level;the expression in senile was significantly higher than that in adulthood and the expression was the lowest in adulthood.The results of immunohistochemistry showed that there was no significant difference in the expression and localization of GCNF in different stages of testicular development,mainly in spermatogenic cells and round sperm cells,and a small amount in interstitial cells and Sertoli cells.By detecting the expression and localization of GCNF in different developmental stages of yak testis,it is found that GCNF may be involved in spermatogenesis,differentiation and deformation,which provides a theoretical basis and new basis for yak reproduction.

关 键 词：生殖细胞核因子 睾丸 生殖 牦牛:圆形精子 表达 定位 

分 类 号：S852.21[农业科学—基础兽医学]