Mouse embryonic stem cells express the cardiac myosin heavy chain genes during development in vitro.
Mouse embryonic stem cells express the cardiac myosin heavy chain genes during development in vitro.
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DOI：
10.1016/s0021-9258(19)38485-6
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发表时间：
1990-07
期刊：
影响因子：
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通讯作者：
Jeffrey Robbins;J. Gulick;Alejandro Sánchez;P. Howles;Thomas Doetschman
中科院分区：
文献类型：
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作者：
Jeffrey Robbins;J. Gulick;Alejandro Sánchez;P. Howles;Thomas Doetschman
在小鼠胚胎中，早期器官发生的特点是形成功能性心肌，因此，9天的胚胎虽然没有完全发育心脏，但已显示出跳动。根据这一观察结果，我们发现有趣的是，小鼠胚状体（EB）可以在体外由全潜能胚胎干细胞发育而来，经历自发的收缩活动。为了确定这些细胞是否能够再现心脏发生的各个方面，我们从跳动的EB中制备了一个cDNA文库，并用鸡骨肌球蛋白重链cDNA进行筛选。我们发现文库中主要的肌球蛋白转录本编码α -和β -心脏亚型。此外，还分离到了一段胚胎骨骼肌球蛋白cDNA。在EB和9天胚胎心脏中发现肌球蛋白重链转录本是相同的。制备转录特异性引物，对单个EB进行聚合酶链反应分析。数据显示，单个EB能够同时表达α -和β -亚型以及极少量的胚胎骨骼转录物。这些数据表明，EB转录合适的组织和发育阶段特异性肌球蛋白重链基因，因此可以作为在分子水平上研究早期心脏生成过程的模型系统。
In the mouse embryo, early organogenesis is characterized by the formation of a functional cardiac muscle, such that 9-day embryos exhibit beating, although not fully developed hearts. In light of this observation, we found it intriguing that mouse embryoid bodies (EB), which can develop in vitro from totipotential embryonic stem cells, undergo spontaneous contractile activity. To determine if these cells are capable of recapitulating aspects of cardiogenesis, a cDNA library was prepared from beating EB and screened with a chicken skeletal myosin heavy chain cDNA. We found that the predominant myosin transcripts in the library encode the alpha- and beta-cardiac isoforms. In addition, an embryonic skeletal myosin cDNA was isolated. The myosin heavy chain transcripts in both EB and 9-day embryonic hearts were found to be the same. Transcript-specific primers were prepared, and polymerase chain reaction analyses on single EB were carried out. The data show that a single EB is capable of expressing both the alpha- and beta-isoforms as well as very low amounts of the embryonic skeletal transcript. These data indicate that EB transcribe the appropriate tissue- and developmental stage-specific myosin heavy chain genes and therefore serve as a model system for studying early cardiogenic processes at the molecular level.