Molecular cloning and characterization of HGF family members with neurotrophic activities

(1) HGF和c-Met/HGF受体在神经系统中的表达分析采用定量竞争RT-PCR和免疫组化检测HGF和c-Met的表达。(a)大脑皮层发育；HGF和c-Met均受到动态调节。结合体外HGF加速存活、神经突和迁移的活性，提示HGF可能在皮层形成中起关键作用。(b)视网膜发育；c-Met和HGF在视网膜细胞发育过程中均有表达。尤其是c-Met在睁眼后视网膜神经元和成人视网膜感光细胞中的表达，提示HGF在视网膜中的活性依赖性作用。(2)体外培养HGF的神经营养活性。观察不同类型神经细胞的存活、神经突延伸、早期神经上皮细胞的增殖和促进迁移活性等神经营养活性。(3) hgf样蛋白（HLP）作为一种新型神经营养因子的鉴定。我们首先用鸡外周神经节细胞确定HLP是一种神经营养因子。(4)利用转基因方法研究HGF在运动神经退行性疾病中的作用(a)在神经元特异性启动子控制下产生过表达HGF的转基因小鼠（NS-HGF-Tg）。对生成的小鼠进行表征。(b) hgf - als转基因小鼠的双转基因小鼠的产生和表征。通过将HS-HGF-Tg与als转基因小鼠配对，与双转基因小鼠、单转基因小鼠或野生姐妹型小鼠进行比较。

(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.