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In vivo monitoring of cardiomyocyte proliferation to identify chemical modifiers of heart regeneration

机译:体内监测心肌细胞增殖以鉴定心脏再生的化学修饰剂

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Adult mammalian cardiomyocytes have little capacity to proliferate in response to injury, a deficiency that underlies the poor regenerative ability of human hearts after myocardial infarction. By contrast, zebrafish regenerate heart muscle after trauma by inducing proliferation of spared cardiomyocytes, providing a model for identifying manipulations that block or enhance these events. Although direct genetic or chemical screens of heart regeneration in adult zebrafish present several challenges, zebrafish embryos are ideal for high-throughput screening. Here, to visualize cardiomyocyte proliferation events in live zebrafish embryos, we generated transgenic zebrafish lines that employ fluorescent ubiquitylation-based cell cycle indicator (FUCCI) technology. We then performed a chemical screen and identified several small molecules that increase or reduce cardiomyocyte proliferation during heart development. These compounds act via Hedgehog, Insulin-like growth factor or Transforming growth factor β signaling pathways. Direct examination of heart regeneration after mechanical or genetic ablation injuries indicated that these pathways are activated in regenerating cardiomyocytes and that they can be pharmacologically manipulated to inhibit or enhance cardiomyocyte proliferation during adult heart regeneration. Our findings describe a new screening system that identifies molecules and pathways with the potential to modify heart regeneration.
机译:成年哺乳动物心肌细胞对损伤的反应增殖能力很弱,这一缺陷是心肌梗死后人心脏再生能力差的基础。相比之下,斑马鱼通过诱导多余的心肌细胞增殖,在创伤后使心肌再生,从而为识别阻止或增强这些事件的操纵提供了模型。尽管成年斑马鱼心脏再生的直接遗传或化学筛选提出了许多挑战,但斑马鱼胚胎是高通量筛选的理想选择。在这里,为了可视化活斑马鱼胚胎中的心肌细胞增殖事件,我们生成了转基因斑马鱼品系,这些品系采用基于荧光泛素化的细胞周期指示器(FUCCI)技术。然后,我们进行了化学筛选,确定了在心脏发育过程中增加或减少心肌细胞增殖的几个小分子。这些化合物通过刺猬,胰岛素样生长因子或转化生长因子β信号传导途径起作用。机械或遗传消融损伤后对心脏再生的直接检查表明,这些途径在再生心肌细胞中被激活,可以在成人成年心脏再生过程中进行药理操作以抑制或增强心肌细胞的增殖。我们的发现描述了一种新的筛选系统,该系统可以识别具有修饰心脏再生潜能的分子和途径。

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