ALTERED TAU CONFORMATION AS AN EARLY MARKER IN AD

本课题主要研究阿尔茨海默病和阿尔茨海默病的纤维组织病理的形成。 它的主要成分，tau蛋白。我们假设进步主义 Tau蛋白的修饰和聚合成直链和成对 螺旋丝(PHF)是由构象变化引起的 使用特定的单抗进行鉴定和量化。 构象选择性抗体，其中大部分是在 阿尔伯特·爱因斯坦医学院彼得·戴维斯的实验室，可以 用于确定作为函数的细胞骨架异常的演变 阿尔茨海默病在易受伤害的大脑区域的进展，并将其与 Tau的结构特征。使用这些抗体，病毒的稳定性 可以对PHF或AD选择性的tau构象进行评估和量化 通过对重组单体tau的亲和力测量，合成 聚合牛磺酸和正宗的PHF牛磺酸。我们认为，tau的早期变化 磷酸化和/或构象导致纤维的形成 AD的病理改变，这些变化是稳定的，可以观察到 在疾病过程中非常早期使用早期标志物单抗 在PHF形成开始之前。这一假设将被检验为 1.我们将在体外模拟tau的结构特征， 作为PHF选择性抗体原位结合的基础。这将是 通过寡核苷酸定向突变和亲和力完成 测量。既依赖磷酸化又依赖构象 抗体将被定性。这一信息将表明 Tau构象依赖性抗体的研究进展 特色化的。这一信息将提示tau的进展。 与纤维形成相关的构象变化 2.我们建议选择新的构象敏感的单抗 分别在AIMS 1和AIMS 3中分析和使用tau抗体；3.我们 将探索tau蛋白发生的早期构象变化 在疾病进展过程中使用定量免疫组织化学-和 在光和EM水平上的细胞化学；以及，4。我们建议确认 原位记录的染色是由构象改变的tau引起的 两种部位捕获ELISA在老年人脑组织易损区的应用 对照人群分为轻、中、重度AD。这些研究将是 与项目一起执行，以将 基底前脑甘丙素过度惰化的纤维病理 胆碱能神经元和特异性神经生长因子受体的表达。数据将是 进一步与MRI测量的结构性萎缩和 行为症状学和电生理测量 功能。

This project focuses on the formation of the fibrillar pathology in AD and on its chief component, tau protein. We hypothesize that progressive modification and polymerization of tau proteins into straight and paired helical filaments (PHFs) result from conformational changes that can be identified and quantified using specific monoclonal antibodies. Conformation-selective antibodies, most of which were produced in the laboratory of Peter Davies at Albert Einstein College of medicine, can be used to determine th evolution of cytoskeletal abnormalities as a function of AD progression in vulnerable brain regions, and to correlate these with structural features of tau. Using these antibodies, the stability of the PHF- or AD-selective tau conformations can be assessed and quantified through affinity measurements against recombinant monomeric tau, synthetic polymeric tau, and authentic PHF tau. We propose that early changes in tau phosphorylation and /or conformation lead to the formation of fibrillar pathology in AD, and that these changes are stabilized and can be observed very early in the disease process using early marker monoclonal antibodies prior to the onset of PHF formation. This hypothesis will be tested as follows; 1. We will model the structural features of tau in vitro that underlie binding of PHF-selective antibodies in situ. This will be accomplished through oligonucleotide-directed mutagenesis and affinity measurements. Both phosphorylation-dependent and conformation-dependent antibodies will be characterized. This information will suggest a progression for tau conformation-dependent antibodies will be characterized. This information will suggest a progression for tau conformational changes associated with the formation of the fibrillar pathology; 2. We propose to select novel monoclonal conformation-sensitive tau antibodies for analysis and use in Aims 1 and 3, respectively; 3. We will probe for early conformational changes in tau protein that occur during disease progression using quantitative immunohisto- and cytochemistry at the light and E.M. level; and, 4. We propose to confirm that the staining documented in situ is due to conformationally altered tau by using two site capture ELISAs in vulnerable brain regions in aged control populations, mild, moderate and severe AD. These studies will be performed in conjunction with projects to correlate the formation of the fibrillar pathology with galanin hyperinervation of the basal forebrain cholinergic neurons and expression of specific NGF receptors. Data will be further correlated with structural atrophy as measured by MRI and behavioral symptomatology as well as electrophysiological measures of function.