CEREBROVASCULAR SIGNAL PATHWAYS IN AGING AND ALZHEIMER'S

本研究的长期目标是识别和表征 脑功能、生化和分子异常 微循环在衰老和痴呆中的作用血脑检查 屏障功能对于理解 神经元细胞死亡在老化和阿尔茨海默病，因为 脑内皮细胞主动调节神经元离子和营养物质 微环境虽然血脑的程度和重要性 衰老和阿尔茨海默病中的屏障异常， 有争议的，大量的证据表明， 血脑屏障 我们假设 异常 微血管受体介导的信号通路可能有助于 阿尔茨海默病患者血脑屏障反应性的改变 BBB的缺失可以是年龄-和/或阿尔茨海默病- 相关.因此，阿尔茨海默氏症患者的微血管反应性 年龄匹配的非痴呆老年患者和i例成人对照中的疾病 患者将接受评估。此外，在年轻人和老年人中进行的实验 啮齿动物是必要的一个直接和现成的材料来源， 可以评估哪些方法、存储和年龄相关的变化。 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，在基础和激动剂下 刺激条件，将从上述组中进行评估， 以确定年龄和疾病相关的变化。实验 计划分析，（1）使用放射性配体的受体结合参数 结合技术，受体连接的Gs功能通过重建， 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测定腺苷酸环化酶水平 积累和[3 H]毛喉素结合，特异性和活性，这 酶通过磷蛋白分析，mRNA表达通过RNA印迹 分析;和（3）蛋白激酶C活性，基础分布和 激动剂刺激的易位，通过蛋白质印迹测定的同种型水平，和 通过北方分析确定消息级别。 结果将定义 细胞内信号通路的功能和调节 血脑屏障，并可能提供一个基础的发展 改善脑血管疾病的合理治疗策略， 阿尔茨海默病的神经元功能

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.