Crucial transcription factors in endoderm and embryonic gut development are expressed in gut-like structures from mouse ES cells
Crucial transcription factors in endoderm and embryonic gut development are expressed in gut-like structures from mouse ES cells
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DOI：
10.1634/stemcells.2005-0344
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发表时间：
2006-03-01
期刊：
影响因子：
5.2
通讯作者：
Torihashi, Shigeko
中科院分区：
文献类型：
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作者：
Matsuura, Rie;Kogo, Hiroshi;Torihashi, Shigeko
小鼠胚胎干（ES）细胞是多能的，并保留形成类似于肠道的器官的潜力，在体外显示自发收缩。这些结构的形态特征及其形成，使用悬滴法产生胚状体（EB）进行评估，似乎与体内相似。为了确定是否有相同的分子机制参与的形成过程中，转录因子的表达模式调节内胚层和肠道发育的小鼠胚胎进行了检查，通过原位杂交和体内表达进行了比较。免疫组化法检测基因产物的表达，双染色法分析表达共定位。结果表明，所有检查的因子，即Sox 17，保留了形成类似于在体外显示自发收缩的肠的器官的潜力。这些结构的形态特征及其形成，使用悬滴法产生胚状体（EB）进行评估，似乎与体内相似。为了确定是否有相同的分子机制参与的形成过程中，转录因子的表达模式调节内胚层和肠道发育的小鼠胚胎进行了检查，通过原位杂交和体内表达进行了比较。免疫组化法检测基因产物的表达，双染色法分析表达共定位。结果表明，所有检测的因子，即Sox 17、Id 2、HNF 3 β/Foxa 2和GATA 4，在Els和肠样结构中均表达。此外，它们的表达模式与小鼠胚胎中的表达模式相似。悬滴期后和长出前的EB已经表达了EB上皮结构中相互共定位的所有因子。这些发现表明，肠状结构的起源是在悬滴期间确定的，肠状结构是在悬滴期间形成的上皮结构。他们还表明，在体外系统中使用小鼠ES细胞模拟体内发育，并应证明内胚层和肠道发育的分子机制的研究是有用的。
Mouse embryonic stem (ES) cells are pluripotent and retain the potential to form an organ similar to the gut showing spontaneous contractions in vitro. The morphological features of these structures and their formation, as assessed using the hanging drop method to produce embryoid bodies (EBs), seem to be similar to those in vivo. To determine whether the same molecular mechanisms are involved in the formation process, the expression pattern of transcription factors regulating endoderm and gut development in the mouse embryo was examined by in situ hybridization and compared with in vivo expression. Expression of gene products was also examined by immunohistochemistry, and expression colocalization was analyzed with double staining. The results showed that all factors examined, that is, Sox17, retain the potential to form an organ similar to the gut showing spontaneous contractions in vitro. The morphological features of these structures and their formation, as assessed using the hanging drop method to produce embryoid bodies (EBs), seem to be similar to those in vivo. To determine whether the same molecular mechanisms are involved in the formation process, the expression pattern of transcription factors regulating endoderm and gut development in the mouse embryo was examined by in situ hybridization and compared with in vivo expression. Expression of gene products was also examined by immunohistochemistry, and expression colocalization was analyzed with double staining. The results showed that all factors examined, that is, Sox17, Id2, HNF3 beta/Foxa2, and GATA4, were expressed in both Ells and gut-like structures. Moreover, their expression patterns were similar to those in the mouse embryo. EBs after the hanging drop period and before outgrowth already expressed all factors that were colocalized with each other in EB epithelial structures. These findings suggest that the origin of the gut-like structure is determined during the hanging drop period and that the gut-like structure is formed as the epithelial structure in EBs during the hanging drop period. They also indicate that the in vitro system using mouse ES cells mimics in vivo development and should prove useful in the study of molecular mechanisms for endoderm and gut development.