MEMBRANE CHANGES IN NEUROLOGIC AND AGING DISEASES

神经系统和衰老疾病中的膜变化

• 批准号: 3120062
• 负责人: MARGUERITE M KAY
• 金额: $ 18.22万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-12 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3120062
• 关键词: Alzheimer's disease; Huntington's disease; affinity chromatography; aging; antibody; antigens; band 3 protein; brain cell; cell death; cerebrospinal fluid; enzyme linked immunosorbent assay; glia; human subject; human tissue; immunochemistry; immunoelectron microscopy; immunoglobulin G; laboratory mouse; laboratory rat; multiple sclerosis; nervous system disorder; neurons; phagocytosis; protein structure function; radiotracer; serum; western blottings

The purpose of this proposal is to investigate the role of band 3 and senescent cell antigen (SCA), an aging antigen that is a degradation product of band 3, in neurological aging and disease. Since SCA is intimately involved in cellular aging and removal, we will elucidate its role in neural aging and disease. We will begin by investigating band 3 structure and function in adult and aging brain. We will determine whether it undergoes the same age related changes as the anion transporter, "band 3," in other cells. The following anion transporter, band 3, changes are observed with cellular aging: increased degradation detected by immunoblotting with band 3 antibodies, decreased anion transport activity (increased Km; decreased Vmax), decreased number of high affinity ankyrin binding sites, and binding of physiologic IgG autoantibodies in situ (1- 27). In addition, the anion transporter, band 3, undergoes an as yet undefined change that results in binding of "980" antibodies to aged band 3. We will quantitate the amount of SCA IgG and amount of SCA on neural tissue of various ages. Kinetic studies will be performed on pieces of perfused brains obtained immediately and at various time intervals following death to determine whether there is an increase in SCA following cell death. Titers of antibodies to band 3, synthetic peptides of band 3 and SCA will be determined with ELISA in both serum and cerebral spinal fluid (CSF) to determine if antibodies are present to promote phagocytosis of neurons by glial cells. Mice will be used for the initial aging studies. However, results will be tested in parallel experiments on human tissue. We will compare the changes in neurological diseases such as Alzheimer's Disease (AD) to those occurring during normal brain aging. We continue working on and characterizing 5 antibodies we have that distinguish between normal human brain tissue and tissue from individuals with AD. The specific aims of the proposed research are: 1. Determine band 3 structural and functional changes during brain aging. 2. Quantitate brain band 3, altered band 3, and aging antigen content during aging. 3. Mechanism of band 3 alterations in aging. 4. Determine whether differences in serum and CSF antibodies to brain band 3 synthetic peptides and SCA are detected between young and old normal individuals (both mice and humans). 5. Determine whether differences in serum and CSF antibodies to brain band 3 peptides and SCA are detected between normal individuals and those with diseases such as Alzheimer's disease, multiple sclerosis, Huntington's chorea, and age-matched controls. 6. Develop monospecific and monoclonal antibodies to SCA, aged band 3, and breakdown products of band 3 for immunoblots, immunohistochemistry, immunoelectron microscopy, and affinity chromatography. 7. Identify structural and functional alterations in band 3 in neurological diseases such as AD. 8. Quantitate brain band 3, altered band 3, and aging antigen content in AD and other neurological diseases.

该提案的目的是调查频段3和 衰老细胞抗原（SCA），衰老的抗原，是一种降解 在神经衰老和疾病中，带3的产物。 因为SCA是 密切参与细胞衰老和去除，我们将阐明其 在神经衰老和疾病中的作用。 我们将首先调查频段3 成人和衰老大脑的结构和功能。 我们将确定是否 它经历与年龄相关的与阴离子转运蛋白相同的变化，“ band 3，“在其他单元格中。以下阴离子转运蛋白，带3，变化为 观察到细胞衰老：通过 带有3抗的免疫印迹，降低阴离子转运活性 （增加km; vmax减少），高亲和力的数量减少 结合位点以及生理IgG自身抗体的结合原位（1- 27）。 此外，阴离子转运蛋白3带3进行了 未定义的变化导致“ 980”抗体与老化频带结合 3。我们将量化神经上的SCA IgG和SCA的量 各个年龄的组织。 动力学研究将在 立即和各个时间间隔获得灌注大脑 死亡后以确定SCA是否有所增加 细胞死亡。 抗体3的抗体滴度，频带3的合成肽 SCA将在血清和脑脊柱中用ELISA确定 流体（CSF）确定是否存在抗体以促进吞噬作用 神经胶质细胞的神经元。 小鼠将用于初始老化 研究。 但是，将在人类的并行实验中测试结果 组织。 我们将比较神经系统疾病的变化，例如 阿尔茨海默氏病（AD）对正常脑老化期间发生的患者。 我们 继续研究和描述5种抗体，我们有 区分正常的人脑组织和组织与个体 与广告。 拟议研究的具体目的是：1。 带3脑老化期间的结构和功能变化。 2。定量 脑带3，改变带3和衰老期间抗原含量。 3。 衰老中带3改变的机理。 4。确定差异是否 在脑带3合成肽和SCA的血清和CSF抗体中 在年轻人和老年人（小鼠和人类）之间检测到。 5。确定血清和CSF抗体的大脑条带的差异是否存在 在普通人和患有的人之间检测到3个肽和SCA 阿尔茨海默氏病，多发性硬化症，亨廷顿的疾病 舞蹈和年龄匹配的控件。 6。发展单特异性和单克隆 SCA的抗体，老年频带3和频段3的分解产物 免疫印迹，免疫组织化学，免疫电子显微镜和亲和力 色谱法。 7。确定频带的结构和功能改变 3在神经疾病中，例如AD。 8。定量脑带3，改变 Band 3，以及AD和其他神经系统疾病中的抗原含量。