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 许多疾病的显著特征，包括阿尔茨海默氏症和 肌萎缩侧索硬化症。这个项目的长期目标是 破译导致神经细胞凋亡的分子机制。 调查人员特别感兴趣的是， 氧化应激导致细胞死亡。这里提出的具体研究 来源于使用PC12细胞和交感神经元培养的实验 作为探索氧化应激引起神经元的方式的模型 死亡。对这些途径的机制的了解将使 特定疗法的设计。申请者将对比和比较两个 凋亡细胞死亡的不同范式：自由基介导的细胞 死亡和营养因子戒断在同一神经元中介导的细胞死亡 细胞类型、PC12细胞和交感神经元。通过利用相同的 神经细胞研究这两条途径它们消除了可能性 路径上的差异是不同的可获得性的结果 促进死亡的分子。他们提出了这样的假设：硝酸盐 氧化物和白介素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，具体目的如下： 1.确定NO如何介导自由基诱导的神经细胞死亡： A.哪些一氧化氮合酶是细胞死亡所必需的？B.NO是否诱导P53 活动。自由基致细胞死亡过程中PARP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种？2.至 确定IL-1B是如何介导细胞死亡的 SOD1.在营养因子剥夺中没有不良影响 范例。A.IL-1B是否增加PC12细胞中的一氧化氮合酶？B.什么是IL-1 PC12细胞上的受体水平？它们在细胞死亡过程中受到调控吗？C.是 IL-1受体基因敲除小鼠的神经元保护免受氧化应激？ VASOD1后bcl2基因转导的PC12细胞中IL-1B是否升高 治疗？这些研究将加深我们对NO和IL-1B如何 在神经细胞的凋亡性死亡中起作用。通过破译细胞凋亡 我们将对疾病的治疗有更好的了解 这就是细胞的异常死亡。

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.