Purkinje cells originate from cerebellar ventricular zone progenitors positive for Neph3 and E-cadherin
Purkinje cells originate from cerebellar ventricular zone progenitors positive for Neph3 and E-cadherin
复制标题
DOI：
10.1016/j.ydbio.2009.11.032
复制
发表时间：
2010-02-15
影响因子：
2.7
通讯作者：
Ono, Yuichi
中科院分区：
文献类型：
--
作者：
Mizuhara, Eri;Minaki, Yasuko;Ono, Yuichi
GABA能浦肯野细胞（PC）提供来自小脑皮质的主要输出，小脑皮质控制运动和姿势。虽然PC的分化机制已经得到了很好的研究，但PC的确切起源和初始特化机制仍有待澄清。在这里，我们确定了小脑和脊髓GABA能祖细胞选择性表面标记，Neph 3，这是一个直接下游的目标基因Ptf 1a，GABA能神经元发育的重要调节。用流式细胞仪从胚胎小脑中分选出Neph 3（+）GABA能祖细胞，并通过体外培养对该群体的细胞命运进行定位。我们发现，大多数从小鼠E12.5小脑分选的Neph 3（+）细胞群体注定要分化成PC，而剩余的一小部分Neph 3（+）细胞是Pax 2（+）中间神经元的祖细胞，Pax 2（+）中间神经元可能是小脑深核GABA能神经元。这些结果通过使用表达Neph 3启动子驱动的GFP的转基因小鼠的短期体内谱系追踪实验得到证实。此外，我们还鉴定了E-cadherin作为选择性表达的标记物，其由位于小脑背侧的Neph 3（+）细胞亚群表达。分选实验显示，胚胎小脑中的Neph 3（+）E-cadherin（高）群体定义了PC祖细胞，而Pax 2（+）中间神经元的祖细胞富集在Neph 3（+）E-cadherin（低）群体中。两者合计，我们的研究结果确定了两个空间划界的亚区，产生不同的小脑GABA能亚型，并揭示了起源的PC在小脑原基的脑室区。(C)2009 Elsevier Inc. All rights reserved.
GABAergic Purkinje cells (PCs) provide the primary output from the cerebellar cortex, which controls movement and posture. Although the mechanisms of PC differentiation have been well studied, the precise origin and initial specification mechanism of PCs remain to be clarified. Here, we identified a cerebellar and spinal cord GABAergic progenitor-selective cell surface marker, Neph3, which is a direct downstream target gene of Ptf1a, an essential regulator of GABAergic neuron development. Using FACS, Neph3(+) GABAergic progenitors were sorted from the embryonic cerebellum, and the cell fate of this population was mapped by culturing in vitro. We found that most of the Neph3(+) populations sorted from the mouse E12.5 cerebellum were fated to differentiate into PCs while the remaining small fraction of Neph3(+) cells were progenitors for Pax2(+) interneurons, which are likely to be deep cerebellar nuclei GABAergic neurons. These results were confirmed by short-term in vivo lineage-tracing experiments using transgenic mice expressing Neph3 promoter-driven GFP. In addition, we identified E-cadherin as a marker selectively expressed by a dorsally localized subset of cerebellar Neph3(+) cells. Sorting experiments revealed that the Neph3(+) E-cadherin(high) population in the embryonic cerebellum defined PC progenitors while progenitors for Pax2(+) interneurons were enriched in the Neph3(+) E-cadherin(low) population. Taken together, our results identify two spatially demarcated subregions that generate distinct cerebellar GABAergic subtypes and reveal the origin of PCs in the ventricular zone of the cerebellar primordium. (C) 2009 Elsevier Inc. All rights reserved.