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
党文庆 1何敏 1曲玉涵 1张凯 1杨小峰 2秦小伟 3韩琦 1郭翔宇 1吕丽华1
作者信息
- 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
- 折叠
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(frizzled 1,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、D114、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(cyclin-dependent 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 11 subfamily 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细胞中,并在OD卵泡颗粒细胞中表达量极显著降低(P＜0.01);两条信号通路中CTNNB1、LEF1、Notch2和Hey2基因的表达量均在OD卵泡颗粒细胞中极显著升高(P＜0.01).本结果表明,牛第1卵泡波偏差过程中F1、F2卵泡颗粒细胞基因表达具有特异性,Wnt/β-catenin和Notch信号通路关键组分的表达趋势一致,这将为探索卵泡发育过程中颗粒细胞功能的分子调控机制提供参考依据.
Abstract
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 factor 1(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).The mRNA expression of CTNNB1,LEF1,Notch2 and Hey2 were all extremely significantly increased in OD follicular granulosa cells(P＜0.01).This study showed that the gene expression in Fl and F2 follicular granulosa cells during the deviation of bovine first follicular wave was specific,and the gene expression of key components in Wnt/β-catenin and Notch signaling pathway had the same trend,which provides a reference basis for the molecular regulatory mechanism of granulosa cell function during follicular development.
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Key words
Bovine follicular granulosa cells/Wnt and Notch signaling pathway/Cell cycle/Apoptosis/Steroid hormone引用本文复制引用
基金项目
山西省基础研究计划项目面上项目(20210302123393)
山西省大同市现代产业研究技术体系项目(农业)(2021027)
山西省基础研究计划项目(202303021221095)
山西农业大学"引进人才科研启动工程"项目(2023BQ56)
出版年
2024
国家科技期刊平台
NSTL
万方数据