Differences in neurogenic potential in floor plate cells along an anteroposterior location: midbrain dopaminergic neurons originate from mesencephalic floor plate cells
Differences in neurogenic potential in floor plate cells along an anteroposterior location: midbrain dopaminergic neurons originate from mesencephalic floor plate cells
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DOI：
10.1242/dev.02879
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发表时间：
2007-09-01
期刊：
影响因子：
4.6
通讯作者：
Imai, Toshio
中科院分区：
文献类型：
--
作者：
Ono, Yuichi;Nakatani, Tomoya;Imai, Toshio
干细胞定向分化纯化中脑多巴胺能神经元是实现安全有效的帕金森病细胞移植治疗的关键问题。虽然最近的研究已经确定了调控MesDA神经元发育的因素，但其背后的机制还没有完全了解。最近，有研究表明，中脑底板(FP)细胞具有神经前体特性，从而产生MesDA神经元。在这里，我们直接通过使用带有FP细胞特异性表面标记的荧光激活细胞分类(FACS)的命运映射实验来检验这一点，并证明了中脑FP细胞具有神经生成活性并在体外生成MesDA神经元。相比之下，分离的尾侧FP细胞并不像之前认为的那样具有神经生成潜力。对携带Lmx1a基因突变的Dreher突变小鼠和在尾部FP细胞中异位表达OTX2的转基因小鼠的分析表明，OTX2决定了赋予FP细胞神经生成活性的前身份，并至少部分通过诱导Lmx1a决定了MesDA的命运。我们进一步证明，FACS可以从胚胎干细胞来源的神经细胞中分离出MesDA前体细胞，这是一种合适的移植材料。我们的数据提供了对MesDA神经元的指定和生成机制的见解，并说明了一种治疗帕金森病的有用的细胞替代方法。
Directed differentiation and purification of mesencephalic dopaminergic (mesDA) neurons from stem cells are crucial issues for realizing safe and efficient cell transplantation therapies for Parkinson's disease. Although recent studies have identified the factors that regulate mesDA neuron development, the mechanisms underlying mesDA neuron specification are not fully understood. Recently, it has been suggested that mesencephalic floor plate (FP) cells acquire neural progenitor characteristics to generate mesDA neurons. Here, we directly examined this in a fate mapping experiment using fluorescence-activated cell sorting (FACS) with an FP cell-specific surface marker, and demonstrate that mesencephalic FP cells have neurogenic activity and generate mesDA neurons in vitro. By contrast, sorted caudal FP cells have no neurogenic potential, as previously thought. Analysis of dreher mutant mice carrying a mutation in the Lmx1a locus and transgenic mice ectopically expressing Otx2 in caudal FP cells demonstrated that Otx2 determines anterior identity that confers neurogenic activity to FP cells and specifies a mesDA fate, at least in part through the induction of Lmx1a. We further show that FACS can isolate mesDA progenitors, a suitable transplantation material, from embryonic stem cell-derived neural cells. Our data provide insights into the mechanisms of specification and generation of mesDA neurons, and illustrate a useful cell replacement approach for Parkinson's disease.