Analysis of differentiation system in cerebral GABAergic neuron and oligodendrocyte progenitors.

脑 GABA 能神经元和少突胶质细胞祖细胞分化系统分析。

基本信息

批准号：
20700291
负责人：
ESUMI Shigeyuki
金额：
$ 2.75万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2009
项目状态：
已结题

项目摘要

GABAergic neurons and oligodendrocyte share many characters in the forebrain development. Both cell types have been reported to originate in the medial ganglionic eminence and migrate to the neocortex. Previously, it has been reported that GABAergic neuron progenitors express not only of GABAergic neuron markers but also ones of oligodendrocytes progenitors, such as NG2 and CNP (Belachew et al., 2003 ; Aguirre et al., 2004 ; Dayer, et al., 2005). Moreover, NG2/Olig2 positive glial progenitors generate both oligodendrocyte and astrocyte in the developing forebrain using NG2creBAC transgenic mouse (Zhu et al., 2008) and Olig2-CreER knock-in mouse (Ono et al., 2008). These previous data raise a possibility that some GABAergic neuron progenitors produce glial cells in the developing forebrain. To test this possibility, here we have performed immunohistochemistry, single-cell RT-PCR, single-cell microarray analysis and immunocytochemistry analyses with glial markers in GAD67-GFP positive cells from GAD67-GFP knock-in mouse brain. As a result, we found that the neuronal markers and glial markers are co-localized in the GAD67-GFP-positive cells at mRNA and protein level. To further investigate cell lineage(s) from GABAergic neuron progenitors in vivo and in vitro, we utilized GAD67-Cre knock-in mice and Z/EG reporter mice. As a result, we found that the neuronal markers and glial markers are co-localized in the GAD67-GFP-positive cells at mRNA and protein level. Finally, to investigate GAD67 lineage in vivo and in vitro, we utilized GAD67-Cre knock-in mice and Z/EG reporter mice. We observed the most of recombined GFP positive cells were GABAergic neuron, but a few cells were oligodendrocyte and astrocyte. At present, we can not completely rule out the possibility that leaky or weak expression of GAD67 gene occur in the small subset of glial progenitors during cell-type specification.

GABA能神经元和少突胶质细胞在前脑发育中具有许多角色。据报道，两种细胞类型均起源于内侧神经节的显着性并迁移到新皮层。以前，据报道，GABA能神经元的祖细胞不仅表达了GABA能神经元标记物，还表达少突胶质细胞祖细胞的祖细胞（例如NG2和CNP）（Belachew等，2003; Aguirre等，2004; Dayer等，2004; Dayer等，2005; Aguirre et al。）。此外，NG2/Olig2阳性神经胶质祖细胞使用NG2CREBAC转基因小鼠（Zhu等，2008）和Olig2-Creer敲入小鼠在发育前脑中产生少突胶质细胞和星形胶质细胞（Ono等，2008）。这些先前的数据提出了一些GABA能神经元祖细胞在发育中的前脑中产生神经胶质细胞的可能性。为了测试这种可能性，我们在这里进行了免疫组织化学，单细胞RT-PCR，单细胞微阵列分析和免疫细胞化学分析，并在GAD67-GFP阳性细胞中使用GAD67-GFP敲击小鼠脑的GAD67-GFP阳性细胞进行了胶质标记。结果，我们发现在mRNA和蛋白质水平的GAD67-GFP阳性细胞中将神经元标记和神经胶质标记物共定位。为了进一步研究体内GABA能神经元祖细胞和体外的细胞谱系，我们利用了GAD67-CRE敲门小鼠和Z/EG报告基因小鼠。结果，我们发现在mRNA和蛋白质水平的GAD67-GFP阳性细胞中将神经元标记和神经胶质标记物共定位。最后，为了研究体内和体外的GAD67谱系，我们使用了GAD67-CRE敲入小鼠和Z/EG报告基因小鼠。我们观察到大部分重组的GFP阳性细胞是GABA能神经元，但一些细胞是少突胶质细胞和星形胶质细胞。目前，我们无法完全排除GAD67基因在细胞类型规范期间的胶质祖细胞中漏水或弱表达的可能性。