Changes in the Expression of Pyruvate Dehydrogenase Kinase Isoenzymes During Early Differentiation of Mouse Embryonic Stem Cells

Saime Güzel
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Abstract

The embryonic stem cells (ESCs) are pluripotent, self-renewing cells that able to differentiate into any of the germ layers involved in embryogenesis. However, the molecular mechanisms that control ESC pluripotency and differentiation remain poorly understood. The family of Pyruvate dehydrogenase kinase (PDK1-4), inactivates the mitochondrial pyruvate dehydrogenase complex via phosphorylation, plays a crucial role in the control of glucose homeostasis. In the current study, gene expression levels of PDK isoenzymes were analyzed on undifferentiated mouse embryonic stem cells (mESCs) and compared to mESCs induced to differentiate by removal of leukemia inhibitory factor (LIF) for 1, 3 and 5 days. Besides, we performed gene expression analysis of several genes related to pluripotency and differentiation. In addition, we also determined glucose uptake rates by a colorimetric assay kit in early differentiated and undifferentiated mESCs. Differently expression level of PDK isoenzymes in pluripotent and LIF-depleted mESCs suggest that they may have roles in differentiation and pluripotency of ESCs. Furthermore, this study lays the foundation for detailed investigation of molecular mechanisms underlying the roles of PDKs in the pluripotency and transition to differentiated state of ESCs.
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丙酮酸脱氢酶激酶同工酶在小鼠胚胎干细胞早期分化过程中的表达变化
胚胎干细胞(ESCs)是一种多能、自我更新的细胞,能够分化成参与胚胎发生的任何胚层。然而,控制ESC多能性和分化的分子机制仍然知之甚少。丙酮酸脱氢酶激酶(PDK1-4)家族通过磷酸化使线粒体丙酮酸脱氢酶复合物失活,在葡萄糖稳态控制中起着至关重要的作用。本研究分析了未分化小鼠胚胎干细胞(mESCs)中PDK同工酶的基因表达水平,并与去除白血病抑制因子(LIF)诱导分化的mESCs进行了1、3和5天的比较。此外,我们还对多能性和分化相关的几个基因进行了基因表达分析。此外,我们还通过比色测定试剂盒测定了早期分化和未分化mESCs的葡萄糖摄取率。PDK同工酶在多能性mESCs和liff缺失mESCs中的不同表达水平表明它们可能在ESCs的分化和多能性中发挥作用。此外,本研究为深入研究PDKs在ESCs多能性和向分化状态转变过程中的分子机制奠定了基础。
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来源期刊
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1
审稿时长
3 months
期刊介绍: The International Journal of Applied Research in Veterinary Medicine promotes excellence in the clinical practice of veterinary medicine by disseminating fundamental scientific, diagnostic, and treatment knowledge gained from prospective and retrospective research in a timely manner. The Journal fulfills its mission through rapid peer review of each submitted article, and publication of all articles within 90 days of acceptance. All published articles meet the standards of Balance, Independence, Objectivity and Scientific Rigor.
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