权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Examination of the number of spinal motor neurons by using selective elimination of motor neurons

通过选择性消除运动神经元检查脊髓运动神经元的数量

基本信息

批准号：
17590167
负责人：
SATO Noboru
金额：
$ 2.24万
依托单位：
Fukushima Medical University
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-17590167/
关键词：
Bax Chick Cell death Neuron Electroporation Doxycycline Rho

项目摘要

To clear how the number of spinal motor neurons is controlled during development (neurogenesis and elimination by cell death), selective elimination of motor neurons has been experimentally developed in this study. Bax, a pro-apoptotic member of the Bcl-2 family, was examined as executioner for motor neurons. To introduce Bax specifically into motor neurons during the period of early PCD, we have used in ovo electroporation coupled with the tet regulatory system. The reverse tetracycline-controlled transactivator was expressed under the control of the motor neuron specific HB9 promoter to target gene expression to spinal motor neurons. When gain-of-function Bax mutants were introduced into chick cervical motor neurons by this method, the number of dying motor neurons was significantly increased compared with those of control chick embryos. Naive Bax did not affect motor neuron survival by this method but significantly induced a large amount of cell death as soon as expressed in the spinal cord using the constitutive promoter such as the CMV promoter for their expression. Taken together, these strategies examined in this study will allow spatially and temporally controlled elimination of spinal motor neurons and will give specific insight into how the number of spinal motor neurons is regulated during development.

为了明确脊髓运动神经元的数量在发育过程中是如何控制的（神经发生和通过细胞死亡消除），在本研究中实验性地开发了运动神经元的选择性消除。Bax是Bcl-2家族的促凋亡成员，被检测为运动神经元的刽子手。为了在早期PCD期间将Bax特异性地引入运动神经元，我们使用了卵内电穿孔结合泰特调节系统。在运动神经元特异性HB 9启动子的控制下表达反向四环素控制的反式激活因子，以将基因表达靶向脊髓运动神经元。当通过这种方法将功能获得性Bax突变体导入鸡颈运动神经元时，与对照鸡胚相比，死亡运动神经元的数量显着增加。通过这种方法，幼稚Bax不影响运动神经元存活，但是一旦在脊髓中表达，使用组成型启动子如CMV启动子用于其表达，就显著诱导大量细胞死亡。总之，在这项研究中检查的这些策略将允许空间和时间控制消除脊髓运动神经元，并将提供具体的见解，脊髓运动神经元的数量是如何在发展过程中进行调节。