CEREBROVASCULAR SIGNAL PATHWAYS IN AGING AND ALZHEIMER'S

衰老和阿尔茨海默病中的脑血管信号通路

• 批准号: 3417346
• 负责人: PAULA GRAMMAS
• 金额: $ 17.63万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 1995-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3417346
• 关键词: Alzheimer's disease; G protein; adenylate cyclase; adrenergic receptor; aging; autoradiography; biological signal transduction; blood brain barrier; cell death; cerebrovascular disorder diagnosis; cerebrovascular disorders; enzyme mechanism; human tissue; laboratory rat; membrane reconstitution /synthesis; messenger RNA; microcirculation; northern blottings; postmortem; protein kinase C; radiotracer; receptor binding; receptor mediated endocytosis; western blottings

The long-term goal of this study is to identify and characterize functional, biochemical and molecular abnormalities of the cerebral microcirculation in aging and dementia. Examination of blood-brain barrier function is critical to understanding the pathogenesis of neuronal cell death in aging and Alzheimer's Disease because the cerebral endothelium actively regulates the neuronal ionic and nutrient microenvironment. Although the extent and importance of blood-brain barrier abnormalities in aging and Alzheimer's disease are controversial, considerable evidence exists for a dysfunctional blood-brain barrier. It is our hypothesis that abnormalities in microvascular receptor-mediated signaling pathways may contribute to an altered blood-brain barrier responsiveness in Alzheimer's disease. Abnormalities of the BBB may be age- and/or Alzheimer's disease disease- related. Consequently, microvascular responsiveness in Alzheimer's disease in age-matched, non-demented elderly patientsand i adult control patients will be assessed. In addition, experiments in young and aged rodents are necessary an an immediate and ready source of material on which methods, storage, and age-related changes can be evaluated. Specifically, microvessel adrenergic and cholinergic receptors, adenylate cyclase, and protein kinase C, under both basal and agonist stimulated conditions, will be assessed from the above groups in order to identify- age and disease-related alterations. Experiments are planned to analyze, (1) receptor binding parameters using radioligand binding techniques, receptor-linked Gs function via reconstitution in S49 cyc (G protein deficient) cells, and levels of Gs and Gi using toxin ribosylation; (2) adenylate cyclase levels by measuring cAMP accumulation and [3H]forskolin binding, specificity and activity of this enzyme by phosphoprotein analysis, and mRNA expression by RNA blot analysis; and (3) protein kinase C activity, basal distribution and agonist-stimulated translocation, isoform levels by Western blots, and message levels by Northern analysis. The results will define the function and regulation of intracellular signalling pathways at the blood-brain barrier and may provide a basis for the development of rational therapeutic strategies for improved cerebrovascular and neuronal function in Alzheimer's disease.

这项研究的长期目标是识别和表征 脑功能性，生化和分子异常 衰老和痴呆中的微循环。检查血脑筋 屏障功能对于理解的发病机理至关重要 衰老和阿尔茨海默氏病的神经元细胞死亡，因为 脑内皮积极调节神经元离子和营养素 微环境。虽然血脑的程度和重要性 衰老和阿尔茨海默氏病的障碍异常是 有争议的证据证明了功能失调 血脑屏障。 我们的假设是 微血管受体介导的信号通路可能有助于 阿尔茨海默氏病的血脑屏障反应改变了。 BBB异常可能是年龄和/或阿尔茨海默氏病 - 有关的。因此，阿尔茨海默氏症的微血管反应能力 年龄匹配的，未痴呆的老年患者和成人对照的疾病 将评估患者。另外，在年轻人和老年的实验 啮齿动物是必要的直接材料来源 可以评估哪些方法，存储和与年龄相关的方法。 具体而言，微血管肾上腺素能和胆碱能受体， 基底和激动剂下的腺苷酸环化酶和蛋白激酶C 刺激条件将从上述组中评估 识别与年龄和疾病相关的改变。实验是 计划分析，（1）使用辐射仪的受体结合参数 结合技术，通过重组的受体连接GS功能 S49 CYC（G蛋白缺乏）细胞以及使用毒素的GS和GI水平 核糖基化； （2）通过测量cAMP来测量腺苷酸环化酶水平 积累和[3H]福斯科蛋白结合，特异性和活性 通过磷蛋白分析和RNA印迹通过磷蛋白分析和mRNA表达的酶 分析; （3）蛋白激酶C活性，基础分布和 激动剂刺激的易位，通过蛋白质印迹的同工型水平，以及 北方分析的消息水平。 结果将定义 细胞内信号通路的功能和调节 血脑屏障，可能为发展提供基础 改善脑血管和的理性治疗策略 阿尔茨海默氏病的神经元功能。