MST1和LATS1在牦牛黄体形成、维持和退化中的表达及HCG对其表达的影响  

The Expression of MST1 and LATS1 in the Formation,Maintenance and Degeneration of Corpus Luteum in Yak(Bos grunniens)and the Effect of HCG on Its Expression

作　　者：周应聪 张同享 王靖雷 赵凌 张瑞 姚颖 钱文洁 崔燕[2,3] 余四九[2,3] 樊江峰[2,3] ZHOU Ying-Cong;ZHANG Tong-Xiang;WANG Jing-Lei;ZHAO Ling;ZHANG Rui;YAO Ying;QIAN Wen-Jie;CUI Yan;YU Si-Jiu;FAN Jiang-Feng(Alxa Zuoqi Animal Disease Prevention and Control Center,Alxa Zuoqi 750300,China;College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep&Goat,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]阿拉善左旗动物疫病预防控制中心,阿拉善左旗750300 [2]甘肃农业大学动物医学院,兰州730070 [3]甘肃农业大学甘肃省牛羊胚胎工程技术研究中心,兰州730070

出　　处：《农业生物技术学报》2024年第8期1844-1856,共13页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31660732);甘肃省科技重点研发计划(农业类)项目(20YF8NA131);陇原青年创新创业人才项目(2021LQGR47);甘肃农业大学科技创新基金(青年导师基金)项目(GAU-QDFC-2018-10);甘肃省高校基本科研业务费。

摘　　要：哺乳动物STe20样激酶1(mammalian STE20-like kinase 1,MST1)和大肿瘤抑制蛋白1(large tumor suppressor kinase 1,LATS1)是Hippo通路核心激酶级联反应链的关键成员,参与雌性动物生殖过程并发挥重要作用。为探究MST1和LATS1与牦牛(Bos grunniens)黄体(corpus luteum,CL)形成、维持和退化之间的关系。本研究采集雌性牦牛黄体组织,按照黄体形成(红体期和有腔黄体期)、维持(妊娠黄体期)和退化(退化黄体期)的不同阶段将材料分为4组,通过qPCR和Western blot检测MST1和LATS1 mRNA及蛋白在黄体形成、维持和退化中的表达差异性,运用免疫组织化学(immunohistochemistry,IHC)对MST1和LATS1在黄体形成、维持和退化中进行定位分析。另外,分离培养牦牛卵巢颗粒细胞(granule cells,GC),通过添加人绒毛膜促性腺激素(human chorionic gonadotropin,HCG)诱导颗粒细胞黄体化,采用qPCR、Western blot和免疫荧光(immunofluorescence,IF)等方法对MST1和LATS1 m RNA及蛋白在颗粒细胞黄体化前后进行定量和定位分析。结果显示,MST1和LATS1 mRNA及蛋白在牦牛黄体不同发育阶段表达量存在差异,在妊娠黄体期MST1表达量最高,而LATS1在有腔黄体期的表达最为丰富,在退化黄体期MST1和LATS1表达量均最低。IHC结果显示,在牦牛粒性黄体细胞中MST1和LATS1分布广泛,且MST1在血管内皮细胞中特征表达。添加5 IU/mL HCG诱导颗粒细胞黄体化后,MST1和LATS1 m RNA及蛋白的表达量极显著上升。通过IF染色发现,MST1和LATS1蛋白主要定位于颗粒细胞细胞质,且黄体化前后位置没有改变。研究结果表明,MST1和LATS1介导Hippo信号通路通过控制颗粒细胞分化,黄体细胞增殖和凋亡,参与雌性牦牛黄体组织的形成、维持、退化以及功能运转,并且与黄体生理过程中各类生殖激素密切相关。本研究为进一步探索Hippo信号通路对牦牛黄体组织大小调控以及调节哺乳动物生殖生理过程的作用机制提供了基Mammalian STe20-like kinase 1(MST1)and large tumor suppressor protein 1(LATS1)are key members of the core kinase cascade of the Hippo pathway,which participate in the reproductive process of female animals and play an important role.This study aimed to explore the relationship between MST1,LATS1,and the formation,maintenance,and degeneration of yak(Bos grunniens)corpus luteum(CL).In this study,the corpus luteum of female yaks was collected and divided into 4 groups according to the different stages of corpus luteum formation(corpus rubrum stage,lacunose luteum stage),maintenance(pregnancy luteal phase)and degeneration(degenerative luteal phase).qPCR and Western blot were used to detect the differences of MST1,LATS1 mRNA and protein expression in the formation,maintenance,and degeneration of the corpus luteum.Immunohistochemistry(IHC)was used to analyze the localization of MST1 and LATS1 in the formation,maintenance,and degeneration of the corpus luteum.In addition,granulosa cells(GC)of the yak ovary were isolated and cultured,and luteinization of granulosa cells was induced by adding human chorionic gonadotropin(HCG).qPCR,Western blot and IF methods were used to analyze the quantity and localization of MST1,LATS1 mRNA and protein before and after luteinization.The results showed that MST1,LATS1 mRNA and protein were expressed in the yak corpus luteum,and there were differences in the expression of MST1 at different developmental stages of corpus luteum.The expression of MST1 was the highest in the pregnancy luteal phase,while the expression of LATS1 was the most abundant in the lacunose luteum stage,and the expression of MST1 and LATS1 was the lowest in the degenerative luteal phase.The results of IHC showed that MST1 and LATS1 were widely distributed in yak granular luteal cells,and MST1 was characterized by expression in vascular endothelial cells.After luteinization of granulosa cells induced by 5 IU/mL HCG,the expression of MST1,LATS1 mRNA and protein increased significantly.By IF staining,it was found that M

关 键 词：牦牛 黄体 哺乳动物STe20样激酶1(MST1) 大肿瘤抑制蛋白1(LATS1) Hippo通路 HCG 

分 类 号：S857.2[农业科学—临床兽医学]