Molecular cloning and characterization of HGF family members with neurotrophic activities

具有神经营养活性的 HGF 家族成员的分子克隆和表征

基本信息

批准号：
10680717
负责人：
FUNAKOSHI Hiroshi
金额：
$ 2.11万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680717/
关键词：
Hepatocyte growth factor HGF c-Met HLP neurotrophic factor neuron Transgenic mice neurodegenerative disease クローニングトランスジェニック Sky Gas6 チロシンキナーゼ型受容体

项目摘要

(1) Expressional analysis of HGF and c-Met/HGF receptor in the nervous systemExpression of HGF and c-Met were examined by quantitative competitive RT-PCR and immunohitochemistry.(a) Cerebral cortex development; both HGF and c-Met was regulated in a dynamic fashion. Taken together with in vitro activities of HGF that accelerate survival, neurite, and migration, it is suggested that HGF may have critical role for the cortical layer formation.(b) Retinal development; both c-Met and HGF were expressed in retinal cells during development. Especially c-Met expression and in retinal neurons after eye opening and adult photoreceptor cells in adult, suggesting the activity dependent role of HGF in retina.(2) Neurotrophic activities of HGF in vitro. Neurotrophic activities, such as survival, neurite extension, proliferation in early neuroepithelial cells and migration promoting activities in different types of neural cells were examined.(3) Identification of HGF-like protein (HLP) as a novel neurotrophic factor. We first identify HLP as a neurotrophic factor using chicken peripheral ganglion cells.(4) Role of HGF on motor neurodegenerative disease using transgenic approach(a) Generation of HGF-overexpressing transgenic mice (NS-HGF-Tg) under the control of neuronal specific promoter. Generated mice were characterized.(b) Generation and characterization of double-transgenic mice of HGF-ALS-transgenic mice. By mating HS-HGF-Tg with ALS-transgenic mice, comparing with double-transgenic mice, single-transgenic mice or sister wild-type mice.

(1)神经系统中HGF和c-Met/HGF受体的表达分析采用定量竞争性RT-PCR和免疫组化方法检测HGF和c-Met的表达。 HGF 和 c-Met 均以动态方式受到调节。结合 HGF 加速存活、神经突和迁移的体外活性，表明 HGF 可能对皮质层形成具有关键作用。(b) 视网膜发育； c-Met 和 HGF 在发育过程中均在视网膜细胞中表达。尤其是睁眼后视网膜神经元和成体感光细胞中c-Met的表达，提示HGF在视网膜中的活性依赖性作用。(2)HGF体外神经营养活性。检测了神经营养活性，如早期神经上皮细胞的存活、神经突延伸、增殖以及不同类型神经细胞的迁移促进活性。(3)鉴定HGF样蛋白(HLP)作为一种新型神经营养因子。我们首先使用鸡外周神经节细胞将HLP鉴定为神经营养因子。(4)使用转基因方法HGF在运动神经退行性疾病中的作用(a)在神经元特异性控制下产生HGF过表达转基因小鼠(NS-HGF-Tg)发起人。对产生的小鼠进行表征。(b)HGF-ALS转基因小鼠双转基因小鼠的产生和表征。通过将HS-HGF-Tg与ALS转基因小鼠交配，与双转基因小鼠、单转基因小鼠或姊妹野生型小鼠进行比较。