牛卵泡颗粒细胞中Wnt/β-catenin与Notch信号通路关键因子的表达规律  

Expression Patterns of Key Factors of Wnt/β-catenin and Notch Signaling Pathway in Bovine(Bos taurus)Follicular Granulosa Cells

作　　者：党文庆[1] 何敏 曲玉涵 张凯 杨小峰 秦小伟 韩琦 郭翔宇 吕丽华 DANG Wen-Qing;HE Min;QU Yu-Han;ZHANG Kai;YANG Xiao-Feng;QIN Xiao-Wei;HAN Qi;GUO Xiang-Yu;LYU Li-Hua(College of Animal Science,Shanxi Agricultural University,Taigu 030801,China;Department of Biology,Xinzhou Normal College,Xinzhou 034000,China;Department of Reproductive Medicine,Shanxi Provincial People's Hospital,Taiyuan 030012,China)

机构地区：[1]山西农业大学动物科学学院,太谷030801 [2]忻州师范学院生物系,忻州034000 [3]山西省人民医院生殖医学科,太原030012

出　　处：《农业生物技术学报》2024年第11期2552-2563,共12页Journal of Agricultural Biotechnology

基　　金：山西省基础研究计划项目(面上项目,20210302123393);山西省基础研究计划项目(202303021221095);山西省大同市现代产业研究技术体系项目(农业,2021027);山西农业大学“引进人才科研启动工程”项目(2023BQ56)。

摘　　要：颗粒细胞的增殖、凋亡和类固醇激素的合成在卵泡的发育过程中发挥重要作用,这些功能受信号通路的调控。本研究分别对牛(Bos taurus)第1卵泡波偏差前(pre-deviation,PD)和偏差发生时(onset of diameter deviation,OD)第1大卵泡F1和第2大卵泡F2的颗粒细胞进行转录组测序并对其中的Wnt/β-catenin与Notch通路的关键组分进行表达量验证,进而探讨两条信号通路、细胞周期和细胞凋亡的关键基因及类固醇激素合成相关基因的表达规律。结果显示,PDF1、PDF2、ODF1和ODF2颗粒细胞中表达量高的通路关键组分对应的基因为:无翼型MMTV整合位点家族成员10a(wingless-type MMTV integration site family,member 10a,Wnt10a)、Wnt2b、卷曲蛋白3(frizzled1,Fzd3)、低密度脂蛋白相关受体8(low density lipoprotein-related receptor 8,LRP8)、蓬乱蛋白3(dishevelled3,Dvl3)、轴蛋白1(Axin1)、糖原合成酶激酶3β(glycogen synthase 3β,GSK3β)、β-catenin(catenin beta 1,CTNNB1)、转录因子7样2(transcription factor 7-like 2,TCF7L2)、淋巴增强因子1(lymphoid enhancer factor1,LEF1)、B-细胞淋巴瘤因子9(B-cell CLL/lymphoma 9,BCL9)及Notch2、Dll4、与YRPW基序蛋白2相关的发状分裂相关增强子(Hes related with YRPW motif2,Hey2);细胞周期相关基因有:细胞周期蛋白2(cyclin2,CCND2)、细胞周期蛋白依赖性激酶4(cyclin-dependent kinase4,CDK4)、细胞周期蛋白依赖性激酶抑制剂1B(cyclindependent kinase inhibitor 1A,CDKN1B);线粒体凋亡途径主要由BCL2-associated X(BCL2-associated X,Bax)基因进行调控,并由Caspase3和Caspase6来执行;类固醇激素合成基因有:3β-羟基类固醇脱氢酶基因(3β-hydroxysteroid dehydrogenase,3β-HSD)、细胞色素P450家族成员11A1(cytochrome P450 family 11subfamily A member 1,CYP11A1)和CYP19A1,且OD卵泡颗粒细胞E2合成能力增强;Wnt2定位于偏差过程的卵泡颗粒细胞中,在OD卵泡颗粒细胞中表达量显著升高(P<0.05);Wnt2b蛋白定位于卵泡颗粒细胞和膜�The proliferation,apoptosis and steroid hormone synthesis of granulosa cells play important roles in follicle development,and these functions are regulated by signaling pathways.In this study,the transcriptome sequencing of granulosa cells from the first large follicle F1 and the second large follicle F2 in bovine(Bos taurus)first follicle wave pre-deviation(PD)and onset of diameter deviation(OD)were separately performed to obtain the expression patterns of genes in Wnt/β-catenin and Notch signaling pathways,cell cycle,apoptosis and steroid hormone synthesis,and to validate the expression of key components of the 2 pathways.The results showed that the genes of key components of the 2 pathways with high expression in PDF1,PDF2,ODF1 and ODF2 follicular granulosa cells were wingless-type MMTV integration site family,member 10a(Wnt10a),Wnt2b,frizzled3(Fzd3),low density lipoprotein-related receptor 8(LRP8),dishevelled 3(Dvl3),Axin1,glycogen synthase 3β(GSK3β),catenin beta 1(CTNNB1),transcription factor 7-like 2(TCF7L2),lymphoid enhancer factor1(LEF1),B-cell CLL/Lymphoma 9(BCL9),Notch2,Dll4 and Hes related with YRPW motif2(Hey2).Cell cycle-related genes with high expression were cyclin2(CCND2),cyclindependent kinase4(CDK4),cyclin-dependent kinase inhibitor 1A(CDKN1A),CDKN1B.The apoptosis of granulosa cells was mainly regulated by BCL2-associated X(Bax)in the mitochondrial apoptosis pathway and executed by Caspase3 and Caspase6.The expression of the steroid genes were 3β-hydroxysteroid dehydrogenase(3β-HSD),cytochrome P450 family 11 subfamily A member 1(CYP11A1),and CYP19A1 were elevated,and E2 synthesis ability of OD follicular granulosa cells was increased compared with that of PD follicles.Wnt2 protein was localized in follicular granulosa cells in deviation process and its expression was significantly higher in OD follicular granulosa cells(P<0.05);Wnt2b protein was localized in follicular granulosa cells and theca cells and its expression was extremely significant lower in OD follicular granulosa cells(P<0.01)

关 键 词：牛卵泡颗粒细胞 Wnt、Notch信号通路 细胞周期 凋亡 类固醇激素 

分 类 号：S823.91[农业科学—畜牧学]