Exhaustive detection and analysis of gene expression associated with radial migration and differentiation in developing neocortical neurons

为了用EGFP标记皮质板细胞，我们在胚胎第13天（E13）对ICR小鼠胚胎进行了子宫内电穿孔。EGFP标记的细胞沿径向向脑膜表面迁移（径向迁移）。转染后，在急性脑切片上解剖E15脑室区、E16中间区和E18皮质板。用蛋白酶、木瓜蛋白酶分散解剖切片。用光学细胞分选法从分散的细胞中分离出EGFP阳性细胞。从EGFP阳性细胞中获得的RNA被扩增，然后通过微阵列分析进行详尽的研究。Sema6C和Syntaphilin在中间区表达量是心室区和皮质板的3倍以上。为了在蛋白水平上证实微阵列的结果，我们用免疫组织化学方法观察了Sema6C和Syntaphilin在皮质中的定位。E17小鼠的Sema6C和Syntaphilin定位在中间区。这些结果表明，Sema6C和Syntaphilin在径向迁移细胞的中间区表达增加。由于Sema6C和Syntaphilin与细胞迁移有关，Sema6C和Syntaphilin可能对中间区径向迁移有功能作用。

To label cortical plate cells by EGFP, in utero electroporation was applied to ICR mice embryos in embryonic day 13 (E13). EGFP labeled cells migrated radially to pial surface (radial migration). After the transfection, ventricular zone of E15, intermediate zone of E16 and cortical plate of E18 were dissected from acute brain slices. The dissected slices were dispersed by using protease, papain. EGFP positive cells were isolated from the dispersed cells by means of optical cell sorting methods. The RNA acquired from the EGFP positive cells were amplified and then investigated exhaustively by microarray analysis. Sema6C and Syntaphilin expressed over three fold in intermediate zone than ventricular zone and cortical plate. To confirm the microarray results at protein level, the localization of Sema6C and Syntaphilin in cortex was observed immunohistochemically. Sema6C and Syntaphilin were localized in intermediate zone in E17 mice. These results suggested that the expression of Sema6C and Syntaphilin in radially migrating cells were increased in intermediate zone. As it is known that Sema6C and Syntaphilin associates with cell migration, Sema6C and Syntaphilin could have functional role for radial migration in intermediate zone.