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

详尽检测和分析与发育中的新皮质神经元径向迁移和分化相关的基因表达

• 批准号: 17500207
• 负责人: FURUKAWA Tomonori
• 金额: $ 1.54万
• 依托单位: Hamamatsu University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17500207/
• 关键词: development; brain; migration; microarray; cell sorting; neuron

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.

为了用EGFP标记皮质板细胞，将子宫内电穿孔应用于胚胎13天（E13）的ICR小鼠胚胎。EGFP标记的细胞放射状迁移至软膜表面（放射状迁移）。转染后，取E15脑室区、E16中间区和E18皮质板。用蛋白酶、木瓜蛋白酶对切片进行分散。通过光学细胞分选方法从分散的细胞中分离EGFP阳性细胞。扩增从EGFP阳性细胞获得的RNA，然后通过微阵列分析进行详尽的研究。Sema6C和Syntaphilin在中间带的表达是室带和皮质板的3倍以上。为了在蛋白质水平上证实芯片结果，化学染色观察了Sema6C和Syntaphilin在皮质中的定位。Sema6C和Syntaphilin定位于E17小鼠的中间区。这些结果表明，Sema6C和Syntaphilin在放射状迁移的细胞中的表达在中间带增加。由于已知Sema6C和Syntaphilin与细胞迁移有关，因此Sema6C和Syntaphilin可能在中间带的放射状迁移中起作用。

项目成果

Transgenic expression of enhanced green fluorescent protein enables direct visualization for physiological studies of vasopressin neurons and isolated nerve terminals of the rat

• DOI: 10.1210/en.2004-0830
• 发表时间: 2005-01-01
• 期刊: ENDOCRINOLOGY
• 影响因子: 4.8
• 作者: Ueta, Y;Fujihara, H;Murphy, D
• 通讯作者: Murphy, D

能動的Cl-ホメオスタシス仮説と実験的てんかん病態モデル

主动氯稳态假说和实验性癫痫病理模型

• 发表时间: 2006
• 期刊: てんかん研究 24
• 作者: Inoue;K. et al.;福田敦夫
• 通讯作者: 福田敦夫

Molecular basis for the GABAA receptor-mediated tonic inhibition in rat somatosensory cortex

• DOI: 10.1093/cercor/bhl087
• 发表时间: 2007-08-01
• 期刊: CEREBRAL CORTEX
• 影响因子: 3.7
• 作者: Yamada, Junko;Furukawa, Tomonori;Fukuda, Atsuo
• 通讯作者: Fukuda, Atsuo

Characterization of newly cloned variant of rat glycine receptor α1 subunit.

新克隆的大鼠甘氨酸受体 α1 亚基变体的表征。

• 发表时间: 2005
• 期刊: Biochem.Biophys.Res.Commun. 327
• 作者: Ban;T.et al.;Toyoda H. et al.;Ueta Y.et al.;Inoue K. et al.
• 通讯作者: Inoue K. et al.

Insertion of α7 nicotinic receptors at neocortical layer V GABAergic synapses is induced by a benzodiazepine, midazolam

苯二氮卓类药物咪达唑仑诱导 α7 烟碱受体插入新皮质 V GABA 能突触

• 期刊: Cereb. Cortex (in press)
• 作者: Heck;N. et al.;Yamamoto.S.et al.
• 通讯作者: Yamamoto.S.et al.