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

The role of astrocytes in maintaing reuronal function

星形胶质细胞在维持肾神经功能中的作用

基本信息

批准号：
07044236
负责人：
KUWANO Ryozo
金额：
$ 8.06万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07044236/
关键词：
astrocytes S100 protein neuronal function diphteria toxin Cre-lox transgenic mice gene expression immunohistochemisty GFAP

项目摘要

We focused on the role of astrocytes in maintaining of neuronal functions and attempted to introduce the strategy of cell ablation using a adiphtheia toxin A fragment (DT-A) gene in transgenic mice.1) Prior to create transgenic mice astrocyte specific gene expression was examined in primary culture cells prepared from mouse embryos. In the course of this project we purified several proteins involved in synapse formation from never growth cone-enriched fraction, determined the amino acid sequence, prepared specific antibodies against these peptides and analyzed subcellular localization in the cultured cells.2) Based on spatial-temporal specific gene expression using as appropriate promoter such as S100 or GFAP promoter in astrocyte, the DT-A gene is activated in the restricted cells in the central nervous system. In addition we used cre-lox system for control of the target gene expression. Cre-recombinase mediated site specific recombination occurs on the restricted DNA sequence composed of 34 bp (loxP). DT-A activity was suppressed by insertion of transcription termination signal down stream of murine molony leukemia virus promoter. The DT-A activated by co-transfection with cre-recombinase resulted in cell death.3) Gene trapping provided novel promoters expressed in the central nervous system. We have found the trapped gene related to mental retardation and characterize an affect of the gene expression on maintaining neuronal functions.

本研究旨在探讨星形胶质细胞在维持神经元功能中的作用，并尝试在转基因小鼠中引入应用白喉毒素A片段（DT-A）基因进行细胞消融的策略。1）在建立转基因小鼠之前，先在从小鼠胚胎制备的原代培养细胞中检测星形胶质细胞特异性基因的表达。本研究从神经生长锥富集组分中纯化了几种参与突触形成的蛋白质，测定了其氨基酸序列，制备了特异性抗体，并分析了其在培养细胞中的亚细胞定位。2）基于以S100或GFAP启动子为合适启动子的星形胶质细胞时空特异性基因表达，DT-A基因在中枢神经系统的限制性细胞中被激活。此外，我们还使用了cre-lox系统来控制目的基因的表达。Cre重组酶介导的位点特异性重组发生在由34 bp组成的限制性DNA序列（loxP）上。DT-A活性被小鼠莫洛尼白血病病毒启动子下游转录终止信号的插入所抑制。基因捕获技术提供了在中枢神经系统表达的新型启动子。我们发现了与智力低下相关的基因，并表征了该基因表达对维持神经元功能的影响。