Formation and function of neural networks in the mouse spinal cord dorsal horn

小鼠脊髓背角神经网络的形成和功能

• 批准号: 17300102
• 负责人: SAITO Tetsuichiro
• 金额: $ 9.86万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17300102/
• 关键词: neuron; development & differentiation; neural network; spinal cord; mammal; transgenic mouse; transcription factor; homeobox gene; ホメオボックス遺伝

We have analyzed molecular mechanisms underlying the formation and function of neural networks in the mouse spinal cord dorsal horn. Axon projection of commissural neurons in the spinal cord has been well studied. However, molecules and mechanisms to regulate their differentiation remained largely unknown. We have established a highly efficient gene transfer method based on in vivo elechroporation in the mouse central nervous system for functional analysis of genes. By gain- and loss-of-function analyses using this method, in combination with transgenic and knockout mice, we have revealed that the fate of commissural neurons is determined by the transcriptional cascade from a proneural gene, Math1, to a homeobox gene, Mbh1, and that Mbh1 is required for the differentiation of commissural neurons. Moreover, chromatin immunoprecipitation assays showed that the Math1 protein directly activates expression of Mbh1 through its enhancer. We have curried out microarray analyses to find downstream genes of the cascade using RNAs from the Mbh1-misexpressed spinal cord. Many genes were activated by misexpression of Mbh1. Some of them, such as Netrin and Slit receptors, Doc and Robes, have been known to play pivotal roles in the projection of axons. Their expressions were regulated in a spatiotemporally organized manner by the cascade from Math1 to Mbhl, suggesting that the cascade is essential for the projection of commiecnral axons.

我们分析了小鼠脊髓背角神经网络形成和功能的分子机制。脊髓连合神经元的轴突投射已得到充分研究。然而，调节其分化的分子和机制仍然很大程度上未知。我们建立了一种基于小鼠中枢神经系统体内电穿孔的高效基因转移方法，用于基因功能分析。通过使用这种方法进行功能获得和功能丧失分析，结合转基因和基因敲除小鼠，我们发现连合神经元的命运是由原神经基因 Math1 到同源盒基因 Mbh1 的转录级联决定的，并且 Mbh1 是连合神经元分化所必需的。此外，染色质免疫沉淀分析表明Math1蛋白通过其增强子直接激活Mbh1的表达。我们已经使用 Mbh1 错误表达的脊髓中的 RNA 进行了微阵列分析，以找到级联的下游基因。许多基因因 Mbh1 的错误表达而被激活。其中一些受体，如 Netrin 和 Slit 受体、Doc 和 Robes，已知在轴突的投射中发挥关键作用。它们的表达通过从 Math1 到 Mbhl 的级联以时空组织的方式进行调节，这表明级联对于共轴突的投射至关重要。

项目成果

Determination of commissural neuronal fate

连合神经元命运的测定

• 发表时间: 2005
• 期刊: PNE 50
• 作者: Tetsuichiro Saito;Daisuke Kawauchi;Satoru Miyagi;Tetsuichiro Saito;斉藤 哲一郎;Daisuke Kawauchi;Satoru Miyagi;Tetsuichiro Saito;Tetsuichiro Saito;Daisuke Kawauchi;斉藤 哲一郎;Tetsuichiro Saito
• 通讯作者: Tetsuichiro Saito

小脳顆粒細胞の分化におけるホメオボックス遺伝子Mbh1の機能

同源盒基因Mbh1在小脑颗粒细胞分化中的功能

• 发表时间: 2007
• 作者: Masugi-Tokita M;Tarusawa E;Watanabe M;Molnar E;Fujimoto K;Shigemoto R;川内 大輔
• 通讯作者: 川内 大輔

交連神経細胞の運命決定機構

连合神经元命运决定机制

• 发表时间: 2005
• 期刊: 蛋白質核酸酵素 50
• 作者: Tetsuichiro Saito;Daisuke Kawauchi;Satoru Miyagi;Tetsuichiro Saito;斉藤 哲一郎;Daisuke Kawauchi;Satoru Miyagi;Tetsuichiro Saito;Tetsuichiro Saito;Daisuke Kawauchi;斉藤 哲一郎
• 通讯作者: 斉藤 哲一郎

ブレインサイエンスレビュー2008

脑科学评论 2008

• 发表时间: 2008
• 作者: Sarna JT;Marzban H;Watanabe M;Hawkes R;斉藤 哲一郎
• 通讯作者: 斉藤 哲一郎

In vivo electroporation in the embryonic mouse central nervous system

• DOI: 10.1038/nprot.2006.276
• 发表时间: 2006-01-01
• 期刊: NATURE PROTOCOLS
• 影响因子: 14.8
• 作者: Saito, Tetsuichiro