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: 3417347
• 负责人: PAULA GRAMMAS
• 金额: $ 18.57万
• 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/3417347
• 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.

本研究的长期目标是确定和表征 大脑功能、生化和分子异常 衰老和痴呆症中的微循环。血脑检查 屏障功能对于理解疾病的发病机制至关重要 衰老和阿尔茨海默病中的神经元细胞死亡是因为 脑内皮积极调节神经元的离子和营养 微环境。尽管血脑的范围和重要性 衰老和阿尔茨海默病中的屏障异常是 有争议的，大量证据表明存在功能失调 血脑屏障。 我们的假设是异常 微血管受体介导的信号通路可能有助于 阿尔茨海默病中血脑屏障反应性的改变。 BBB 异常可能与年龄和/或阿尔茨海默病有关 有关的。因此，阿尔茨海默病的微血管反应性 年龄匹配、非痴呆老年患者和成人对照的疾病 将对患者进行评估。此外，还针对年轻人和老年人进行了实验 啮齿动物是必要的直接且现成的材料来源 可以评估哪些方法、存储和与年龄相关的变化。 具体来说，微血管肾上腺素能和胆碱能受体， 腺苷酸环化酶和蛋白激酶 C，在基础剂和激动剂下 刺激条件，将从上述组中按顺序进行评估 识别与年龄和疾病相关的变化。实验是 计划分析，(1) 使用放射性配体的受体结合参数 结合技术，通过重建受体连接的 Gs 功能 S49 cyc（G 蛋白缺陷）细胞，以及使用毒素的 Gs 和 Gi 水平 核糖基化； (2)通过测量cAMP来测定腺苷酸环化酶水平 该物质的积累和 [3H]forskolin 结合、特异性和活性 通过磷蛋白分析检测酶，通过 RNA 印迹分析 mRNA 表达 分析; (3) 蛋白激酶 C 活性、基础分布和 激动剂刺激的易位、蛋白质印迹的亚型水平，以及 北方分析的消息级别。 结果将定义 细胞内信号通路的功能和调节 血脑屏障，可能为发育提供基础 改善脑血管的合理治疗策略 阿尔茨海默病中的神经元功能。