【目的】探究蛋白质代谢在鸡雄性生殖细胞分化过程中的作用机制,为完善鸡胚胎干细胞(embryonic stem cell,ESC)体外向雄性生殖细胞诱导分化体系研究提供依据。【方法】采用流式分选的方法获取高纯度的ESC、原始生殖细胞(primordial germ cells,PGCs)和精原干细胞(spermatogonia stem cell,SSCs),分别提取细胞的总RNA,采用RNA-Seq方法对3种细胞的转录本进行深度测序,然后进行WEGO(web gene ontology)和KEGG(kyoto encyclopedia of genes and genomes)通路富集分析,筛选出鸡雄性生殖细胞分化过程中参与蛋白质代谢的关键通路及其关键基因,RT-q PCR(Real time Quantitative PCR)检测部分关键差异基因的表达变化,并与RNA-Seq(RNA sequencing)结果进行比较分析,同时分别从体内和体外水平对关键基因NOS2进行抑制,观察各分组不同天数的细胞形态变化及检测NOS2和C-kit、Cvh、Stra8、Dazl、integrinα6和integrinβ1等...
【英文摘要】
【Objective】 The aim of this study was to explore the regulatory mechanism of protein metabolism during the differentiation process of chicken male germ cells and provide a basis for improving the induction system of chicken embryonic stem cells(ESCs) differentiation to male germ cells in vitro.【Method】RNA sequencing was performed using FACS-sorted cells from ESCs,PGCs(primordial germ cells) and SSCs(spermatogonial stem cells),and enrichment analysis,WEGO(Web Gene ontology) and KEGG(Kyoto Encyclopedia of Gen...
Regulatory Study of Protein Metabolism During theDifferentiation Process of Chicken Male Germ CellsLI Dong,TANG Bei-bei,WANG Ying-jie,JI Yan-qin,WANG Fei,LU Zhen-yu,WANG Man,ZHANG Ya-ni,LI Bi-chun【研究意义】生殖细胞在生命循环和传承中有着不可替代的作用,因
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