AGING AND ALZHEIMER DEMENTIA: ROLE OF FIBROUS PROTEIN

该项目旨在了解生物化学，生物学和 阿尔茨海默病（AD）的分子病理学。 在这种疾病中， 主要的组织病理学损害是神经系统的存在， 神经纤维缠结和神经炎斑 阿尔茨海默氏神经系统 已经发现缠结（ANT）含有独特抗原决定簇， 决定因素共同的几个正常的细胞骨架成分。 的 ANT与神经丝（NF）的关系将进一步研究 通过确定NF-ANT决定因素在 NF分子。 所使用的技术是有限的蛋白水解，化学 NF蛋白的切割和免疫印迹。 正常和AD中的三种蛋白质 分子量为58- 70 kD的脑匀浆最近被证明 与单克隆抗ANT抗体反应。 分派该等 正常和AD脑中的蛋白质及其与ANT积累的关系将 用ELISA和免疫印迹技术进行研究。 两个NF-ANT共用 已经发现抗原决定簇对磷酸酶敏感。 到 为了确定ANT是否由磷酸化蛋白组成，我们将从ANT中纯化ANT， 对死于阿尔茨海默氏症的老年性痴呆的受试者进行尸检 类型. 将研究纯化样品的磷酸盐含量。 在 此外，还将测定它们的钙和铝含量， 在缠结的神经元中高浓度。 金属元素将是 通过能量色散X射线荧光测定。 我们的注意力也将 集中于ANT抗原的形态学和生化特性， 精细的神经突起 电镜和免疫电镜 将用于确定抗原是否分布在丝状 结构类似于PHF。 我们还计划确定携带抗原的 结构与PHF一样耐溶解。 最后一 具体目的是调查啮齿动物与人类的关系 PFH，并研究细胞骨架的变化过程中积累的 动物组织中的PHF。 组织化学，电子显微镜和 将使用免疫电子显微术。

This project is aimed at understanding the biochemistry, biology and molecular pathologenesis of Alzheimer's disease (AD). In this disorder, the main histopathological lesion is the presence of neurofibrillary tangles and neuritic plaques in the cerebrum. Alzheimer's neurofibrillary tangles (ANT) have been found to contain unique antigenic determinants and determinants in common with several normal cytoskeletal components. The relationship between ANT and neurofilaments (NF) will be examined further by identifying the topographical distribution of NF-ANT determinants in the NF molecules. The techniques to be used are limited proteolysis, chemical cleavage of NF proteins and immunoblots. Three proteins in normal and AD brain homogenates of molecular weight 58-70kd have recently been shown to react with monoclonal anti-ANT antibodies. The distribution of these proteins in normal and AD brain and their relation to ANT accumulation will be studied by ELISA and immunoblotting technique. Two NF-ANT common antigenic determinants have been found to be sensitive to phosphatase. To find out if ANT consist of phosphorylated proteins, we will purify ANT from autopsy brains of subjects who died of senile dementia of the Alzheimer type. The purified samples will be studied for phosphate content. In addition they will be assayed for calcium and aluminum which are present at high concentration in tangle bearing neurons. The metal elements will be determined by energy disperse x-ray fluorescence. Our attention will also focus on the morphological and biochemical properties of ANT antigens in fine neuronal processes. Electron microscopy and immunoelectron microscopy will be used to determine if the antigens are distributed in a filamentous structure similar to PHF. We also plan to determine if the antigen bearing structures are as resistant to solubilization as the PHF. The last specific aim is to investigate the relationship between rodent and human PFH, and to study the cytoskeletal changes proceeding the accumulation of PHF in animal tissue. Histochemistry, electron microscopy and immunoelectron microscopy will be used.