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

Mechanism of motoneuron network formation

运动神经元网络形成机制

基本信息

批准号：
14380359
负责人：
TANAKA Hideaki
金额：
$ 9.09万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

We performed cDNA screening by a signal sequence trap method in order to identify signaling molecules expressed by motoneurons. Motoneurons were purified by an immuno-panning method using SC1 monoclonal antibody from E5 chick embryonic spinal cords. We got three function unknown cDNAs, tentatively named 10B4, 273 and 12D3, and produced monoclonal antibodies against all three proteins. 10B4 and 273 are transmembrane proteins and contain leucine rich repeat (LRR) in their extracellular domain. Since the chimeric proteins of 10B4-AP (alkaline phosphatase) and 273-AP did not bind to any embryonic tissues, we expected that they might be receptors of soluble ligands. In fact 10B4 is found to be a homologue of human LINGO-1, which has been reported as a co-receptor of Nogo/p75 receptor complex. Although 273 is transiently expressed in the motoneuron cell body at the early phase of development, it is continuously expressed in the distal axons. Therefore we expect that 273 might play roles of receptor against some factors derived from muscle or Schwann cells. Soluble protein, 12D3, is produced by roof plate cells, migrated and heavily deposited at the dorsolateral basement membrane of the spinal cord.To reveal the functions of these proteins in ovo we performed gain-of-function and loss-of-function analysis by methods of electroporation and siRNA. However, we have not yet found their clear functions, and plan to continue further analysis of these proteins using both chick and zebrafish systems.

为了鉴定运动神经元表达的信号分子，我们采用信号序列陷阱方法进行cDNA筛选。用SC1单克隆抗体对E5鸡胚胎脊髓中的运动神经元进行免疫筛选纯化。我们获得了三个功能未知的cdna，暂定名为10B4、273和12D3，并产生了针对这三个蛋白的单克隆抗体。10B4和273是跨膜蛋白，其胞外结构域含有丰富亮氨酸重复序列（LRR）。由于10B4-AP（碱性磷酸酶）和273-AP的嵌合蛋白不与任何胚胎组织结合，我们推测它们可能是可溶性配体的受体。事实上，10B4被发现是人类LINGO-1的同源物，而LINGO-1已被报道为Nogo/p75受体复合物的共受体。273虽然在发育早期在运动神经元细胞体中短暂表达，但在远端轴突中持续表达。因此，我们推测273可能对来自肌肉或雪旺细胞的某些因子起受体作用。可溶性蛋白12D3由顶板细胞产生，迁移并大量沉积于脊髓背外侧基底膜。为了揭示这些蛋白在卵细胞中的功能，我们通过电穿孔和siRNA的方法进行了功能获得和功能丧失分析。然而，我们还没有发现它们的明确功能，并计划继续使用小鸡和斑马鱼系统对这些蛋白质进行进一步分析。