In vitro analysis of the origin, migratory behavior, and maturation of cortical pyramidal cells
In vitro analysis of the origin, migratory behavior, and maturation of cortical pyramidal cells
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DOI：
10.1002/cne.10421
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发表时间：
2002-12-02
影响因子：
2.5
通讯作者：
Murakami, F
中科院分区：
文献类型：
--
作者：
Hatanaka, Y;Murakami, F
在发育过程中，神经元通过多种机制从它们的起源位置迁移到它们的最终目的地。尽管越来越多的证据表明，来自皮质心室区（VZ）的细胞径向迁移并产生锥体细胞，但将径向迁移细胞的起源和终末表型直接联系起来的证据有限。此外，这些细胞的迁移行为与其成熟形态之间的关系仍然不清楚。为了解决这些问题，我们开发了一种体外制备方法，使来自皮质VZ的细胞可视化。将编码绿色荧光蛋白（GFP）的质粒注入侧脑室，然后电穿孔，标记胚胎期18 ~ 19天的大鼠皮层VZ细胞。然后将皮层切片并进行器官典型培养。1天后，GFP（+）细胞表现出神经祖细胞和放射状胶质细胞的性质。在接下来的几天里，许多GFP（+）细胞向脑脊液表面迁移；将前导过程向头部表面延伸，并留下一个几乎到达VZ的薄拖尾过程。这些神经元的前导突表达微管相关蛋白2阳性，部分神经元在第5天或第6天转化为树突样树突结构，其后突表现出预示轴突的形态特征。延时分析证实了尾随过程的延伸。分子标记的表达和形态学分析表明，绝大多数迁移的GFP（+）细胞分化为锥体细胞样的兴奋性神经元。这些结果有力地表明，来源于皮层VZ的细胞产生的神经元呈放射状迁移。这些神经元似乎在前面延伸前瞻性树突，在后面留下前瞻性轴突，随后分化成锥体细胞。(C) 2002 Wiley-Liss, Inc。
During development neurons migrate from their site of origin to their final destinations under a variety of mechanisms. Although evidence has been accumulating that the cells from cortical ventricular zone (VZ) migrate radially and produce pyramidal cells, evidence that directly links the origin and the terminal phenotype of radially migrating cells has been limited. Further, the relation between the migratory behavior of these cells and their mature morphology remains obscure. To address these issues, we developed an in vitro preparation that enables visualization of cells derived from the cortical VZ. VZ cells of a rat cortex at embryonic days 18 to 19 were labeled by injecting green fluorescent protein (GFP)-encoding plasmid into the lateral ventricle, followed by electroporation. The cortex was then sliced and cultured organotypically. After 1 day, GFP(+) cells exhibited neural progenitor and radial glial cell natures. Over the next few days, many GFP(+) cells migrated toward the pial surface; extending leading processes toward the pial surface and leaving a thin trailing process that almost reached the VZ. The leading processes of these neurons were positive for microtubule-associated protein 2, and some transformed into dendritic arbor-like structures by day 5 or 6, and their trailing processes exhibited morphologic features indicative of prospective axons. Time-lapse analysis confirmed extension of the trailing processes. Expression of molecular markers and morphologic analysis demonstrated that the vast majority of the migrated GFP(+) cells differentiated into excitatory neurons with pyramidal, cell-like morphology. These results strongly suggested that cells derived from the cortical VZ generate neurons that migrate radially. These neurons appeared to extend prospective dendrites in front and leave prospective axons behind, subsequently differentiating into pyramidal cells. (C) 2002 Wiley-Liss, Inc.