NEURONAL DEGENERATION--MECHANISMS AND PREVENTION

描述（改编自申请人的摘要）神经细胞死亡是一种 许多疾病的突出特征，包括阿尔茨海默病和 肌萎缩侧索硬化症 该项目的长期目标是 破译导致神经元细胞凋亡的分子机制。 特别是，调查人员感兴趣的是， 氧化应激导致细胞死亡。 本文提出的具体研究 这是由使用PC 12细胞和交感神经元培养物的实验引起的 作为模型来探测氧化应激引起神经元损伤的方式， 死亡 了解这些途径的机制将有助于 设计具体的治疗方案。 申请人将对比和比较两个 细胞凋亡的不同模式：自由基介导的细胞 死亡和营养因子撤除介导的同一神经元细胞死亡 细胞类型、PC 12细胞和交感神经元。 通过利用相同的 神经元细胞来研究这两种途径， 不同途径的差异是由于不同的 死亡促进分子。 他们提出假设， 氧化物和白细胞介素-1B是自由基介导的死亡所必需的， 发生. 他们将审查这些假设，具体目标如下： 1. 为了确定NO如何介导自由基诱导的神经元细胞死亡： a.哪些NOS种类是细胞死亡所必需的？ B. NO是否诱导p53 活动 C. PARP在自由基诱导的细胞死亡中是否被激活？ D.是 过氧亚硝酸盐是NO介导的细胞死亡的关键物质？ 2. 到 确定IL-1B如何介导由下调 SOD 1在营养因子缺乏时无毒性作用 范例 a. IL-1B是否增加PC 12细胞NOS活性？ B.什么是IL-1 PC 12细胞上的受体水平 它们在细胞死亡中受到调控吗？ C.是 从IL-1受体敲除小鼠的神经元保护氧化应激？ D.转染bcl-2的PC 12细胞在V-ASOD 1后IL-1B是否增加 治疗？ 这些研究将加深我们对NO和IL-1B 在凋亡性神经元细胞死亡中起作用。 通过破译细胞凋亡 死亡途径，我们将有更好的了解疾病的治疗， 这是异常的细胞死亡。

DESCRIPTION (adapted from applicant's abstract) Neuronal cell death is a prominent feature of many diseases, including Alzheimer's disease and amyotrophic lateral sclerosis. The long term goals of this project are to decipher the molecular mechanisms leading to apoptotic neuronal cell death. In particular, the investigators are interested in the mechanisms by which oxidative stress leads to cell death. The specific studies proposed here arise from experiments using cultures of PC12 cells and sympathetic neurons as models to probe the means by which oxidative stress causes neuronal death. An understanding of the mechanisms of these pathways would allow the design of specific therapies. The applicants will contrast and compare two different paradigms of apoptotic cell death: free radical mediated cell death and trophic factor withdrawal mediated cell death in the same neuronal cell types, PC12 cells and sympathetic neurons. By utilizing the same neuronal cells to study these two pathways they eliminate the possibility that differences in the pathways are the result of availability of different death promoting molecules. They have developed the hypotheses that nitric oxide and interleukin-1B are required for free radical mediated death to occur. They will examine these hypotheses with the following specific aims: 1. To determine how NO mediates free radical induced neuronal cell death: a. Which NOS species are necessary for cell death? b. Does NO induce p53 activity. c. Is PARP activated in free radical induced cell death? d. Is peroxynitrite a critical species in NO mediated cell death? 2. To determine how IL-1B mediates cell death induced by the down-regulation of SOD1 while it has no deleterious effects in the trophic factor deprivation paradigm. a. Does IL-1B increase NOS in PC12 cells? b. What are the IL-1 receptor levels on PC12 cells? Are they regulated in cell death? c. Are neurons from IL-1 receptor knockout mice protected from oxidative stress? d. Is IL-1B increased in bcl-2 transfected PC12 cells after V-ASOD1 treatment? These studies will enhance our understanding of how NO and IL-1B operate in apoptotic neuronal cell death. By deciphering the apoptotic death pathway we will have better insight into the treatment of diseases in which there is aberrant cell death.