The positional identity of mouse ES cell-generated neurons is affected by BMP signaling
The positional identity of mouse ES cell-generated neurons is affected by BMP signaling
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DOI：
10.1007/s00018-012-1182-3
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发表时间：
2013-03-01
影响因子：
8
通讯作者：
Cremisi, Federico
中科院分区：
文献类型：
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作者：
Bertacchi, Michele;Pandolfini, Luca;Cremisi, Federico
我们研究了骨形态发生蛋白（BMPs）在确定小鼠胚胎干细胞（ESCs）体外产生的神经元的位置身份的影响，这是迄今为止被忽视的一个方面。在低等脊椎动物中的经典胚胎学研究表明，BMP抑制多能胚胎细胞的默认命运，这是神经和前部的。此外，当在含有BMP抑制剂的基本培养基中培养时，哺乳动物ESC更有效地产生神经元。在本文中，我们表明，小鼠胚胎干细胞产生，分泌，并在体外神经分化过程中对骨形成蛋白作出反应。在基本培养基中神经化后，分化的ESC显示出与中脑身份一致的基因表达谱，如通过分析许多前后和背腹身份的标记物所评估的。我们发现，BMP内源性产生的神经分化过程中，主要通过抑制端脑基因的表达，这是揭示了与头蛋白或其他BMP抑制剂的治疗。为了更好地表征骨形态发生蛋白对位置命运的影响，我们比较了分化的胚胎干细胞与胚胎前脑、中脑和后脑的基因表达谱。头蛋白和视黄酸（RA）支持神经元分化的胚胎干细胞，但他们表现出不同的影响，其位置的身份：而RA支持典型的基因表达谱的后脑神经元，头蛋白诱导的轮廓特征的背端脑神经元。我们的研究结果表明，内源性产生的骨形成蛋白影响的神经元，胚胎干细胞在体外分化时，在基本培养基中自发产生的位置的身份。这些数据也支持存在一个内在的程序与背端脑的身份神经元分化。我们的ESC神经化方法允许通过在体内胚胎发育过程中发现的相同信号快速分化神经细胞，并通过单独抑制BMP获得皮质身份。
We investigated the effects of bone morphogenetic proteins (BMPs) in determining the positional identity of neurons generated in vitro from mouse embryonic stem cells (ESCs), an aspect that has been neglected thus far. Classical embryological studies in lower vertebrates indicate that BMPs inhibit the default fate of pluripotent embryonic cells, which is both neural and anterior. Moreover, mammalian ESCs generate neurons more efficiently when cultured in a minimal medium containing BMP inhibitors. In this paper, we show that mouse ESCs produce, secrete, and respond to BMPs during in vitro neural differentiation. After neuralization in a minimal medium, differentiated ESCs show a gene expression profile consistent with a midbrain identity, as evaluated by the analysis of a number of markers of anterior-posterior and dorsoventral identity. We found that BMPs endogenously produced during neural differentiation mainly act by inhibiting the expression of a telencephalic gene profile, which was revealed by the treatment with Noggin or with other BMP inhibitors. To better characterize the effect of BMPs on positional fate, we compared the global gene expression profiles of differentiated ESCs with those of embryonic forebrain, midbrain, and hindbrain. Both Noggin and retinoic acid (RA) support neuronal differentiation of ESCs, but they show different effects on their positional identity: whereas RA supports the typical gene expression profile of hindbrain neurons, Noggin induces a profile characteristic of dorsal telencephalic neurons. Our findings show that endogenously produced BMPs affect the positional identity of the neurons that ESCs spontaneously generate when differentiating in vitro in a minimal medium. The data also support the existence of an intrinsic program of neuronal differentiation with dorsal telencephalic identity. Our method of ESC neuralization allows for fast differentiation of neural cells via the same signals found during in vivo embryonic development and for the acquisition of cortical identity by the inhibition of BMP alone.