| 摘要: |
| 【目的】探究牛优势卵泡颗粒细胞（GCs）和膜细胞（TCs）中基因的表达差异，筛选并研究与生殖激素合成相关的基因。【方法】在GEO数据库获得GSE83524芯片数据，通过在线软件GEO2R筛选牛优势卵泡中GCs和TCs之间的差异表达基因。使用DAVID软件对获得的差异表达基因进行GO功能富集分析；使用KOBAS软件进行KEGG信号通路分析，筛选出GCs和TCs之间参与生殖激素合成相关信号通路；使用String结合Cytoscape软件构建蛋白与蛋白之间的相互作用网络（PPI）。以牛的优势卵泡为材料，采用Real-time PCR对筛选出的差异表达基因进行验证。【结果】经在线软件GEO2R分析，共获得247个差异表达基因，其中43个表达上调，204个表达下调；247个差异表达基因的GO功能分析结果共分为3大类41组，其中参与生物学过程（BP）的有 22组，参与细胞组分（CC）的有12组，参与分子功能（MF）的有7组；KEGG信号通路分析共发现17条通路，其中腺苷酸环化酶3基因（ADCY3）、细胞色素P450 17A1基因（CYP17A1）、腺苷酸环化酶8基因（ADCY8）、鸟嘌呤核苷酸结合蛋白基因（GNAS）、骨形态发生蛋白6基因（BMP6）5个差异表达基因参与了卵巢类固醇生成通路。经PPI网络互作分析，筛选出了前10位的Hub基因；Real time PCR结果显示，CYP17A1、ADCY8在TCs中的表达量极显著高于GCs (P<0.01)；ADCY3、BMP6在TCs中的表达量显著高于GCs(P<0.05)，GNAS在GCs中的表达量极显著高于TCs(P<0.01)，这5个基因在GCs和TCs中的表达趋势与GEO芯片数据结果一致。【结论】ADCY3、CYP17A1、ADCY8、GNAS、BMP6在GCs与TCs之间表达量存在显著差异，其可能在牛优势卵泡GCs和TCs中对生殖激素的合成及分泌产生了重要作用。 |
| 关键词: 牛 优势卵泡 差异表达基因 颗粒细胞 膜细胞 生殖激素 |
| DOI： |
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| 基金项目:贵州省科技计划项目（黔科合基础［2020］1Y138）；国家自然科学基金青年基金项目（31702097）；铜仁市科技计划项目（铜市科研［2020］80号）；铜仁学院生态畜牧创新团队项目（CXTD［2020-19］） |
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| Screening and analysis of genes related to reproductive hormone synthesis in granulosa cells and theca cells of cattle dominant follicles |
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ZHAO Yuanyuan,WU Zhenyang,AN Qingming,et al
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| Abstract: |
| 【Objective】This study explored differential expressions of genes in granulosa cells (GCs) and theca cells (TCs) of cattle dominant follicles and genes related to reproductive hormone synthesis were screened and studied.【Method】GSE83524 chip data were obtained from GEO database,and the differentially expressed genes between GCs and TCs of dominant follicles in cattle were screened by GEO2R.GO function enrichment was analyzed using DAVID software to the differentially express genes,and KOBAS software was used to perform KEGG signal pathway analysis.Reproductive hormones related signaling pathways were screened from GCs and TCs,and protein protein interaction network (PPI) was constructed through String combined with Cytoscape software. Using bovine dominant follicles as materials,the selected reproductive hormones related genes were verified by real-time PCR.【Result】After analysis by online software GEO2R,247 differentially expressed genes were obtained,among which 43 were up-regulated and 204 were down-regulated.GO function analysis was performed on 247 differentially expressed genes,and they were divided into 41 groups in three categories.There were 22 groups involved in biological process (BP),12 groups involved in cellular component (CC),and 7 groups involved in molecular function (MF).KEGG signaling pathway analysis revealed a total of 17 pathways,among which 5 differentially expressed genes,including ADCY3,CYP17A1,ADCY8,GNAS and BMP6,were involved in the ovarian steroid production pathway.Top 10 Hub genes were screened by PPI network interaction analysis.Real-time PCR results showed that expression levels of CYP17A1 and ADCY8 in TCs were significantly higher than that of GCs (P<0.01).Expression levels of ADCY3 and BMP6 in TCs were extremely significantly higher than that of GCs (P<0.05) and expression level of GNAS in GCs was extremely significantly higher than that of TCs (P<0.01).The expression trend of these five genes in GCs and TCs was consistent with the results of GEO chip data.【Conclusion】There were significant differences in expression levels of ADCY3,CYP17A1,ADCY8,GNAS and BMP6 between GCs and TCs,which may play an important role in synthesis and secretion of reproductive hormones in bovine dominant follicle GCs and TCs. |
| Key words: cattle dominant follicles differentially expressed genes granulosa cells theca cells reproductive hormones |
