Genetically-defined lineage tracing of Nkx2.2-expressing cells in chick spinal cord
Genetically-defined lineage tracing of Nkx2.2-expressing cells in chick spinal cord
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DOI：
10.1016/j.ydbio.2010.10.007
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发表时间：
2011-01-15
影响因子：
2.7
通讯作者：
Ikenaka, Kazuhiro
中科院分区：
文献类型：
--
作者：
Gotoh, Hitoshi;Ono, Katsuhiko;Ikenaka, Kazuhiro
在脊髓中，少突胶质细胞（Oligs）的产生完全依赖于Olig 2的存在，Olig 2是一种碱性螺旋-环-螺旋转录因子。然而，它也需要Nkx2.2来产生，其表达遵循Olig 2的表达。尽管据信少突胶质细胞起源于pMN结构域。Nkx2.2存在于位于pMN结构域腹侧的p3结构域中。根据最近的报道，除了鸡脊髓中的pMN结构域之外，少突胶质细胞还可能直接来源于p3结构域。为了分析脊髓中OL的发育，鸡胚胎被广泛用于通过电穿孔进行遗传修饰或用于移植实验，因为与小鼠胚胎相比，鸡胚胎相对容易操作。然而，通过电穿孔的遗传修饰不适合用于神经胶质发育分析，因为神经胶质在成熟之前剧烈增殖。为了克服这些问题，我们建立了一种将外源基因永久导入特定细胞类型的新方法。我们引入了CAT 1基因，鼠逆转录病毒受体，通过电穿孔，然后注射鼠逆转录病毒。利用此方法，我们成功地将小鼠逆转录病毒导入鸡神经管。我们通过Nkx2.2增强子限制CAT 1表达来分析来自p3结构域的细胞谱系，发现当细胞在cE 4处标记时，大多数标记的细胞变成OL。此外，标记的OL被发现遍及脊髓中的白色物质，包括最背侧脊髓。因此，p3结构域直接在鸡脊髓中产生脊髓OL。(C)2010年爱思唯尔公司All rights reserved.
In the spinal cord, generation of oligodendrocytes (OLs) is totally dependent on the presence of Olig2, a basic helix-loop-helix transcription factor. However, it also requires Nkx2.2 for its generation, whose expression follows the expression of Olig2. Although it is believed that oligodendrocytes originate from the pMN domain. Nkx2.2 is present in the p3 domain located ventral to the pMN domain. According to recent reports, it is possible that oligodendrocytes are directly derived from the p3 domain in addition to the pMN domain in the chick spinal cord. We examined this hypothesis in this paper.To analyze OL development in the spinal cord, chick embryos are widely used for genetic modification by electroporation or for transplantation experiments, because it is relatively easy to manipulate them compared with mouse embryos. However, genetic modification by electroporation is not appropriate for glial development analyses because glia proliferate vigorously before maturation. In order to overcome these problems, we established a novel method to permanently introduce exogenous gene into a specific cell type. We introduced the CAT1 gene, a murine retroviral receptor, by electroporation followed by injection of murine retrovirus. By using this method, we successfully transduced murine retrovirus into the chick neural tube. We analyzed cell lineage from the p3 domain by restricting CAT1 expression by Nkx2.2-enhancer and found that most of the labeled cells became OLs when the cells were labeled at cE4. Moreover, the labeled OLs were found throughout the white matter in the spinal cord including the most dorsal spinal cord. Thus p3 domain directly generates spinal cord OLs in the chick spinal cord. (C) 2010 Elsevier Inc. All rights reserved.