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

阿尔茨海默病（AD）和帕金森病（PD）是最常见的疾病。 神经退行性疾病（NDD）在美国流行， 残疾人是残疾人的主要来源， 个人和社会的巨大代价。的分子原因 在AD和多巴胺中皮质和间脑神经元的进行性死亡 PD中的神经元尚不清楚，也没有有效的药理学 阻止疾病进展的策略。最近开发的 技术（细胞质杂种=“胞质杂种”）允许基因转移 AD的特异性线粒体电子传递链（ETC）缺陷 将血小板线粒体DNA（复合物IV）或PD（复合物I）克隆到细胞中， 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设， 生物能量受损的线粒体。 该提案中的实验利用当代微透析， 分子技术，以解决四个特定的目标，测试四个 关于AD和PD中神经元死亡原因的假说。目标1将 表征脑羟自由基（OH*）的药理学 急性局部抑制线粒体ETC复合物后的产生 I与MPP+或IV与叠氮化物输注或急性淀粉样肽输注。 目的2将表征保护酶系统（过氧化氢酶， 谷胱甘肽过氧化物酶，超氧化物歧化酶）， PD和对照受试者，并将确定胞质杂交体细胞是否暴露于 神经营养因子分子（NGF，BDNF）改变ROS产生和凋亡 暴露于MPP+或叠氮化物后，保护性基因的转录 酶或保护酶的活性。目标3将测量mRNA 用定量RT-PCR和蛋白质印迹法测定蛋白水平 和/或ELISA检测生长调节蛋白p53、bcl-2、bcl-xL和 Bax在AD、PD和对照受试者的胞质杂交细胞中的表达 MPP+叠氮化物暴露过程。Aim 4将探讨bcl-2基因的参与。 2、bcl-xL和bax调节胞质细胞程序性死亡 来源于AD、PD或对照受试者。无论在哪里，细胞的结果将 将其扩展到动物中以提供体外与体内的相关性。 每个具体目标的实验结果都具有潜在的治疗价值 应用程序，并将提供适用于其他人类的知识 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.