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Localization of glial-cell line derived neurotrophic factor receptor (GFRa1) in the brain

胶质细胞源性神经营养因子受体 (GFRa1) 在大脑中的定位

基本信息

批准号：
09670654
负责人：
NAKAMURA Shinichi
金额：
$ 1.79万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

Glial cell line-derived neurotrophic factor (GDNFR) has been shown to have trophic effect upon dopaminergic neurons and motoneurons in the anterior horn cells of spinal cord. In the present research project we have planned to localize GDNFR in the brain immunohistochemically and examine whether GDNFR is involved in the pathophysiology of neurodegenerative disease such as Parkinson's disease, amyotrophic lateral sclerosis, multiple system atrophy and Alzheimer's disease.We have firstly developed an anti-peptide antibody against synthetic peptide of amino-terminal portion of GDNFR The antibody recognized a single protein of molecular weight of 80 kDa in the rat brain homogenate. The preabsorption of the antibody with the immunogen completely abolished the immunostaining in the western blot lane. The immunohistochemical study has demonstrated GDNFR-immunoreactivity exclusively in neurons and no glial cells were immunopositive. The immunopositive neuronal somata were found throughout the rat brain. In the neocortex the positive neurons were distributed in the layer II through to layer VI.Pyramidal and non-pyramidal cell were both labeled. In the basal forebrain, numerous large sized neurons were immunolabeled in the nucleus of Meynert, medial septal nucleus, nucleus, of diagonal band of Broca and ventral pallidum. Numerous neuronal somata were found also in nuclei of the hind brain, including substantia nigra, raphe, and cochlear nucleus. Thus, GDNF responsive neurons appear to be distributed more widespread in the rat brain than previously described.

胶质细胞源性神经营养因子（GDNFR）对脊髓前角细胞内的多巴胺能神经元和运动神经元具有营养作用。在本研究项目中，我们计划用化学方法定位GDNFR在脑中的位置，并检查GDNFR是否参与神经退行性疾病的病理生理学，如帕金森病，肌萎缩侧索硬化症，本实验室首次研制出抗氨基-β-半乳糖苷合成肽的抗肽抗体，GDNFR的末端部分抗体识别大鼠脑匀浆中分子量为80 kDa的单一蛋白质。抗体与免疫原的预吸收完全消除了免疫印迹泳道中的免疫染色。免疫组织化学研究表明，GDNFR免疫反应只在神经元和神经胶质细胞免疫阳性。免疫阳性神经元胞体遍布整个大鼠脑。在新皮层，阳性神经元分布于Ⅱ层至Ⅵ层，锥体细胞和非锥体细胞均被标记。在基底前脑，Meynert核、内侧隔核、Broca斜角带核和腹侧苍白球内有大量大型神经元被免疫标记。在黑质、中缝和耳蜗核等中脑核团中也发现了大量神经元胞体。因此，GDNF反应性神经元似乎比先前描述的在大鼠脑中分布更广泛。