NEURODEGENERATION AND NEUROPROTECTION IN MND

这项提议使用体外和体内方法来探索 运动中活跃的神经退行性变和神经保护机制 神经元病(MND)。有两种主要的疾病情况是 目标：家族性肌萎缩侧索硬化症(FALS)与 铜锌超氧化物歧化酶1型(SOD1)(FALS)(1-3)突变和 与法规中的删除关联的废弃(WST/WST)鼠标 增殖细胞核抗原基因的区域(4)。其中之一 这个项目中的假设是有一条类似的路径 在这两种疾病过程中的神经元死亡。 这个项目的一个主要目标是阐明细胞死亡的机制， 并确定在治疗中重要的药物和治疗方法 关于MND的救援--特别是与SOD1有关的FAL和被浪费的 老鼠。在第一个具体目标中，我们将调查 突变型SOD1s和从突变型SOD1中拯救神经元的识别试剂- 诱导细胞死亡。我们将表达野生型和突变型SOD1基因 在培养和中枢神经系统(CNS)中使用原代神经元 重组复制缺陷型腺病毒(ADV)作为载体，以及 开始确定这种表达对自由基的影响 SOD1的积聚、神经元功能和活性以及聚集。我们 也将在酵母中表达野生型和突变型SOD1基因以阐明 酶的突变是否会导致聚集和 不解性。我们将测试不同特工的营救能力 体外培养的SOD1突变型细胞诱导细胞凋亡，然后检测 这些药物是否会影响FALS转基因小鼠的疾病(在 第三个具体目标)。在第二个具体目标中，我们将调查 废弃小鼠神经元死亡的原因。我们将研究神经元 以测试这些细胞的水平是否降低 增殖细胞核抗原信使，经历(体外和体内)自发凋亡，或 更容易受到氧化应激等应激条件的影响。一个 对FALS中MN死亡的发病机制有更好的认识 废弃小鼠可能阐明散发性肌萎缩侧索硬化症细胞死亡机制(S) 和其他神经退行性过程，也可能导致 确定MNDs的有效治疗方法。

This proposal uses in vitro and in vivo approaches to explore neurodegenerative and neuroprotective mechanisms that are active in motor neuron diseases (MNDs). There are two main disease conditions that are targeted: familial amyotrophic lateral sclerosis (FALS) associated with mutations in CU-Zn superoxide dismutase type 1 (SOD1)(FALS) (1-3) and the wasted (wst/wst) mouse which is associated with a deletion in a regulatory region of the PCNA (proliferating cell nuclear antigen) gene (4). One of the hypothesis in this project is the there is a similar pathway with respect to neuronal death in both of these disease processes. A major goal of this project is to clarify the mechanisms of cell death, and to identify agents and therapeutic approaches that are important in rescue with respect to MND-specifically SOD1 linked FALS and the wasted mouse. In the first specific aim, we will investigate the "toxicity" of mutant SOD1s and identify agents that rescue neurons from mutant SOD1- induced cell death. We will express wild type and mutant SOD1 genes in primary neurons in culture and in the central nervous system (CNS) using recombinant replication-defective adenoviruses (AdVs) as vectors, and begun to determine the effect of this expression on free radical accumulation, neuronal function and viability, and aggregation of SOD1. We will also express wild type and mutant SOD1 genes in yeast to clarify whether a mutation of change in the enzyme leads to aggregation and insolubility. We will test varied agents for their ability to rescue in vitro cultured cells from mutant SOD1-induced apoptosis, and then test whether these agents will affect disease in the FALS transgenic mouse (in the third specific aim). In the second specific aim we will investigate the reasons for neuronal death in the wasted mice. We will study neurons from the wasted mouse to test whether these cells have a decreased level of PCNA message, undergo spontaneous apoptosis (in vitro and in vivo), or are more susceptible to stress conditions such as oxidative stress. A better understanding of the pathogenesis of MN death in FALS and the wasted mouse may clarify the mechanism(s) of cell death in sporadic ALS and other neurodegenerative processes and also potentially lead to the identification of effective treatments for MNDs.