A1+3.CA+2 AND MITOCHONDRIA IN ALZHEIMER'S DISEASE

这项工作的长期目标是了解分子机制。 人类潜在的阿尔茨海默病(AD)和年龄相关变化 大脑。许多研究表明，观察到的变化 大脑老化和一些神经退行性疾病，包括阿尔茨海默病，可能 可能是由于高血压性脑脊髓炎引起的胞浆游离CA+2水平异常 A1+3的水平。建议是A1+3或其衍生物之一可以 争夺线粒体上的钙结合部位导致钙离子的释放 进入细胞质。这项初步研究的目的是为了获得足够的 初步数据，以证明一个全面的建议，以研究影响 线粒体和细胞内钙池高水平的A1+3。作为一名 检测系统，人成纤维细胞来源于家族性和非家族性白血病患者 将使用与正常年龄和性别匹配的散发性AD患者作为对照。 荧光探头和定量视频增强显微镜将 用于确定细胞是否暴露于不同浓度的 A1+3可影响膜相关钙离子和细胞内游离钙离子水平 和/或改变线粒体功能。将使用的探头包括： 膜结合钙离子指示剂氯四环素； 细胞内游离钙离子探针；罗丹明123，其特异性染色 线粒体被认为依赖于线粒体膜 潜力。这些实验的积极结果将提供一个基础 为了以后将这项工作扩展到A1+3对神经细胞的影响 在文化上。这样的研究及其扩展应该会增加我们的 了解A1+3的神经毒性效应，最终可能有助于 线粒体改变和钙离子调节对大鼠脑缺血再灌注损伤的影响 神经系统的细胞。他们也可能有助于提供一种理解 阿尔茨海默病和人类大脑中与年龄相关的变化背后的机制。

The long range aim of the work is to understand the molecular mechanisms underlying Alzheimer's disease (AD) and age-related changes in the human brain. A number of studies have indicated that alterations observed in aging brains and in some neurodegenerative disorders, including AD, could be due to abnormal levels of free cytosolic CA+2 brought about by high levels of A1+3. The suggestion is that A1+3 or one of its derivatives can compete for Ca+2 binding sites on mitochondria causing release of the Ca+2 into the cytosol. The aim of this pilot study is to obtain enough preliminary data to justify a full scale proposal to study the effects of high levels of A1+3 on mitochondria and intracellular Ca+2 pools. As a test system, human fibroblasts derived from patients with familial and sporadic AD and from normal age and sex matched controls will be used. Fluorescent probes and quantitative video intensification microscopy will be used to determine whether exposure of cells to various concentrations of A1+3 can affect levels of membrane-associated and free intracellular Ca+2 and/or alter mitochondrial function. The probes to be used are: chlorotetracycline, an indicator of membrane-associated Ca+2; quin2, a probe for free cytosolic Ca+2; and rhodamine 123 whose specific staining of mitochondria is thought to be dependent upon mitochondrial membrane potential. A positive result from these experiments would provide a basis for a later extension of this work into the effects of A1+3 on neural cells in culture. Such studies and their extensions should increase our understanding of the neurotoxic effects of A1+3 and may eventually serve to delineate the effects of alterations of mitochondria and Ca+2 regulation on cells of the nervous system. They may also help provide an understanding of the mechanisms behind AD and age related changes in the human brain.