FREE RADICALS AND CELL DEATH IN MODELS OF ALZHEIMER'S AND PARKINSON'S DISEASE

阿尔茨海默病(AD)和帕金森氏病(PD)是最常见的 美国流行的神经退行性疾病(NDD)，困扰着几个人 数以百万计的公民，是老年人残疾的主要来源 个人和社会付出了巨大的代价。致病原因的分子机制 阿尔茨海默病患者皮质和间脑神经元进行性死亡与多巴胺 帕金森病中的神经元尚不清楚，也没有有效的药物作用。 阻止这两种疾病发展的策略。一种新近开发的 技术(细胞质杂交体=“胞质杂交体”)允许遗传转移 阿尔茨海默病患者线粒体特异性电子传递链缺陷 (络合物IV)或PD(络合物I)将线粒体DNA血小板转化为克隆体 神经元宿主细胞，表明ETC缺陷的病因是在 线粒体基因组。Cybrid细胞提供了探索的模型系统 关于细胞死亡原因的假说 生物能量受损的线粒体。 该方案中的实验利用当代微渗析和 分子技术解决了四个特定的目标，测试了四个 阿尔茨海默病和帕金森病患者神经元死亡原因的假说。目标1将 脑羟基自由基(OH*)的药理研究 线粒体ETC复合体急性局部抑制后的产生 静注MPP+或静注叠氮或急性淀粉样多肽。 目标2将表征保护性酶系统(过氧化氢酶， 线粒体中的谷胱甘肽过氧化物酶、超氧化物歧化酶)， PD和对照受试者，并将确定环体细胞是否暴露于 神经营养因子分子(NGF、BDNF)改变ROS生成和细胞凋亡 暴露于MPP+或叠氮后，基因转录保护 酶，或保护酶的活性。目标3将测量信使核糖核酸 定量逆转录聚合酶链式反应和蛋白质印迹分析 和/或生长调节蛋白P53、BCL-2、BCL-XL和 阿尔茨海默病患者、帕金森病患者和对照组受试者胞质细胞中Bax的表达 MPP+叠氮暴露疗程。目标4将探讨BCL的参与- 2、bcl-xl和bax在调控胞质细胞程序性死亡中的作用 源自AD、PD或对照受试者。无论在哪里，结果都会导致单元格 可推广到动物体内，以提供体外和体内的相关性。 每个特定目标的实验结果都有潜在的治疗作用。 应用程序，并将提供适用于其他人的知识 神经退行性疾病。

Alzheimer's disease (AD) and Parkinson's Disease (PD) are the most prevalent neurodegenerative diseases (NDD) in America, afflict several million citizens, and represent major sources of disability for the individual and substantial cost to society. The molecular causes of progressive death of cortical and diencephalic neurons in AD and dopamine neurons in PD are not known, nor ar there effective pharmacologic strategies for halting progression of either disease. A recently developed technique (cytoplasmic hybrids= "cybrids") has allowed genetic transfer of specific mitochondrial electron transport chain (ETC) defects from AD (complex IV) or PD (complex I) platelet mitochondrial DNA into clonal neuronal host cells, showing that the etiology of the ETC defects is in the mitochondrial genome. Cybrid cells provide model systems to explore hypotheses about the causes of cell death resulting from having bioenergetically impaired mitochondria. The experiments in this proposal utilize contemporary microdialysis and molecular techniques to address four Specific Aims that test four hypotheses about the causes of neuronal death in AD and PD. Aim 1 will characterize the pharmacology of brain hydroxyl free radical (OH*) production following acute, local inhibition of mitochondrial ETC complex I with MPP+ or IV with azide infusion or acute amyloid peptide infusion. Aim 2 will characterize expression of protective enzyme systems (catalase, glutathione peroxidase, superoxide dismutase) in cybrids derived from AD, PD and control subjects and will determine if exposure of cybrid cells to neurotrophin molecules (NGF, BDNF) alters ROS production and apoptosis following exposure to MPP+ or azide, transcription of genes for protective enzymes, or activities of protective enzymes. Aim 3 will measure mRNA levels with quantitative RT-PCR and protein levels with Western blotting and/or ELISA for the growth regulatory proteins p53, bcl-2, bcl-xL, and Bax in cybrid cells derived from AD, PD and control subjects during the course of MPP+ azide exposure. Aim 4 will explore the involvement of bcl- 2, bcl-xL and bax in regulating programmed cell death in cybrid cells derived from AD, PD, or control subjects. Wherever, results in cells will be extended into animals to provide correlation of in vitro with in vivo. The results of experiments in each Specific Aim have potential treatment applications and will provide knowledge applicable to other human neurodegenerative diseases.