NEURONAL CYTOSKELETAL ALTERATIONS IN ALZHEIMER DISEASE

阿尔茨海默病中的神经细胞骨架改变

• 批准号: 3119465
• 负责人: KHALID IQBAL
• 金额: $ 14.42万
• 依托单位: INSTITUTE FOR BASIC RES IN DEV DISABIL
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-02-01 至 1995-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3119465
• 关键词: Alzheimer's disease; SDS polyacrylamide gel electrophoresis; autoradiography; cytoskeletal proteins; cytoskeleton; electron microscopy; human tissue; laboratory rat; microtubule associated protein; neurochemistry; neurofibrillary tangles; neurons; paired helical filament; phosphorylation; protein kinase; tau proteins; western blottings

The long term objective of tis proposal is to learn the etiology and the pathogenesis of Alzheimer disease/senile dementia of the Alzheimer type (AD/SDAT) which constitutes one of the major public health problems in modern society. In the United States alone presently over three million senior citizens are affected and these numbers will keep increasing at a frightening rate unless the disease is understood and prevented. Several lines of evidence suggest that the state of phosphorylation/ dephosphorylation of neuronal proteins including the microtubule associated proteins tau might be affected in AD/SDAT and that the Alzheimer paired helical filaments (PHF) might contain abnormally phosphorylated tau. The specific aims of this project are 1) a thorough investigation of the ability of Alzheimer brain tau and PHF, with or without prior in vitro dephosphorylation, to stimulate in vitro assembly of microtubules from bovine tubulin-determined by turbidimetric measurements, negative stain electron microscopy and SDS-PAGE; 2) identification of the protein kinase/s responsible for the phosphorylation of tau and PHF in AD/SDAT brain and measurements of this kinase activity in AD/SDAT and in age-matched unaffected brains-- determined by autoradiography and Western blots of SDS-PAGE; and 3) study the effect of phosphorylation on the self assembly of tau into filaments -- determined by negative stain electron microscopy, turbidimetric measurements and SDS-PAGE. The studies will test whether the abnormal phosphorylation of tau might be the cause of the microtubule assembly defect in AD/SDAT brain and whether dephosphorylation of tau/PHF can reverse this defect in vitro. Identification of the protein kinase/s responsible for the phosphorylation of tau in AD/SDAT brain and determination of the conditions for the self assembly of tau into filaments and their depolymerization might be critical to devise a rational approach in correcting this defect. These studies will help eluciate how alterations of the normal cytoskeleton might contribute to neurofibrillary pathology and functional deficits in AD/SDAT brain.

提议的长期目标是学习病因和 阿尔茨海默氏病/老年痴呆症的发病机理 （AD/SDAT）构成了主要公共卫生问题之一 现代社会。 仅在美国，目前就超过300万 老年人受到影响，这些数字将不断增加 令人恐惧的速度，除非理解和预防该疾病。 几条证据表明，磷酸化/ 神经元蛋白的去磷酸化，包括微管 相关蛋白质tau可能会在AD/SDAT中受到影响，并且 阿尔茨海默氏症配对的螺旋丝（PHF）可能含有异常含有 磷酸化的tau。 该项目的具体目的是1）彻底 研究阿尔茨海默氏症脑tau和phf的能力，或 没有事先的体外去磷酸化，以刺激体外组装 牛微管蛋白微管的微管确定 测量，负染色电子显微镜和SDS-PAGE； 2） 蛋白激酶的鉴定 AD/SDAT脑中Tau和PHF的磷酸化以及该测量的磷酸化 AD/SDAT和年龄匹配的未受影响的大脑中的激酶活性 - 由自显影和SDS-PAGE的蛋白质印迹确定； 3） 研究磷酸化对tau自组装的影响 细丝 - 由负染色电子显微镜确定， 浊度测量和SDS-PAGE。 研究将测试tau异常磷酸化是否可能 成为AD/SDAT大脑中微管组装缺陷的原因 tau/phF的去磷酸化是否可以在体外扭转这一缺陷。 蛋白激酶的鉴定 tau在AD/SDAT大脑中的磷酸化和测定 将tau自我组装成细丝的条件及其 解放可能至关重要 纠正此缺陷。 这些研究将有助于阐明 正常细胞骨架的改变可能有助于 AD/SDAT脑中的神经原纤维病理和功能缺陷。