首页> 外文期刊>The Journal of Reproduction and Development >Effects of whole genome duplication on cell size and gene expression in mouse embryonic stem cells
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Effects of whole genome duplication on cell size and gene expression in mouse embryonic stem cells

机译:全基因组复制对小鼠胚胎干细胞大小和基因表达的影响

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摘要

Alterations in ploidy tend to influence cell physiology, which in the long-term, contribute to species adaptation and evolution. Polyploid cells are observed under physiological conditions in the nerve and liver tissues, and in tumorigenic processes. Although tetraploid cells have been studied in mammalian cells, the basic characteristics and alterations caused by whole genome duplication are still poorly understood. The purpose of this study was to acquire basic knowledge about the effect of whole genome duplication on the cell cycle, cell size, and gene expression. Using flow cytometry, we demonstrate that cell cycle subpopulations in mouse tetraploid embryonic stem cells (TESCs) were similar to those in embryonic stem cells (ESCs). We performed smear preparations and flow cytometric analysis to identify cell size alterations. These indicated that the relative cell volume of TESCs was approximately 2.2-2.5 fold that of ESCs. We also investigated the effect of whole genome duplication on the expression of housekeeping and pluripotency marker genes using quantitative real-time PCR with external RNA. We found that the target transcripts were 2.2 times more abundant in TESCs than those in ESCs. This indicated that gene expression and cell volume increased in parallel. Our findings suggest the existence of a homeostatic mechanism controlling the cytoplasmic transcript levels in accordance with genome volume changes caused by whole genome duplication.
机译:倍性的改变倾向于影响细胞生理,从长远来看,它有助于物种的适应和进化。在神经和肝脏组织以及致瘤过程中,在生理条件下观察到多倍体细胞。尽管已经在哺乳动物细胞中研究了四倍体细胞,但是对由全基因组重复引起的基本特征和改变仍然知之甚少。这项研究的目的是获得有关整个基因组复制对细胞周期,细胞大小和基因表达的影响的基础知识。使用流式细胞仪,我们证明了小鼠四倍体胚胎干细胞(TESCs)中的细胞周期亚群类似于胚胎干细胞(ESCs)中的细胞周期亚群。我们进行了涂片准备和流式细胞仪分析,以鉴定细胞大小的改变。这些表明,TESC的相对细胞体积约为ESC的2.2-2.5倍。我们还使用外部RNA定量实时PCR研究了全基因组复制对管家和多能性标记基因表达的影响。我们发现,目标成绩单在TESC中的含量是在ESC中的2.2倍。这表明基因表达和细胞体积平行增加。我们的发现表明存在根据全基因组复制引起的基因组体积变化控制细胞质转录水平的体内平衡机制。

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