NEURONAL PLASTICITY IN ALS SPINAL CORD NEURONS

说明：（申请人摘要）本提案的重点是 研究了与运动神经元相关的因果事件， 肌萎缩性侧索硬化症（ALS）中的变性。 目前的理论 关于ALS运动神经元丧失的发病机制，包括但不限于 仅限于中枢神经系统（CNS）兴奋性毒性、自身免疫性 反应，超氧化物歧化酶基因突变，如果家族性ALS，不适用 中枢或外周营养支持丧失。 有趣的是，这些事件 也被证明参与了与编程相关的过程， 细胞死亡或凋亡。 本申请的第一具体目的 研究ALS中程序性细胞死亡标志物的表达 尸检材料 这将通过检查ALS和对照脊髓 脊髓和运动皮质切片显示DNA片段化， 凋亡的特征。 在第二个具体目标中，我们将使用 原位杂交，在某些情况下，免疫细胞化学，以调查 某些细胞死亡基因及其蛋白产物在ALS中的表达 组织. 第三个具体目标将涉及ALS脑筛查 脑脊液（CSF）中是否存在已知的免疫球蛋白（例如Fas/Apo-1） 参与程序性细胞死亡 这些研究的结果将 确定程序性细胞死亡在ALS中的作用，并提供深入了解 潜在的治疗策略来阻止这种细胞死亡过程。 一 目前正在研究的治疗方法是治疗ALS 有神经营养因子的患者 使用这些分子的基本原理 包括理解神经营养因子促进 神经元的存活和再生过程，并且它们显示出某种程度的 对它们有效的神经元群体的选择性。 如果 营养因子疗法是有效的，这是必要的，以证明 肌萎缩侧索硬化症运动神经元表现出对这种治疗作出反应的能力， 战略 第一步是证明ALS运动神经元表达 营养因子信号转导所必需的受体。 第四 具体目标将研究这些受体的表达， 目前正在研究的营养因子作为治疗剂， 治疗ALS。 这些受体包括trks， 神经营养素的高亲和力受体，以及神经营养素的α亚基。 CNTF受体。 我们将使用原位杂交来研究 ALS尸检材料中编码这些受体的特定mRNA。 在 此外，还应该有足够的神经营养因子受体抗体 蛋白质变得可用，我们将纳入逻辑 免疫细胞化学实验来补充mRNA表达的研究。 总的来说，这些研究旨在促进我们对 ALS中的运动神经元变性，这些运动神经元是否维持 对某些营养因子治疗作出反应的能力，并证明 程序性细胞死亡在这场毁灭性的 神经退行性疾病

DESCRIPTION: (Applicant's Abstract) The focus of this proposal is to examined the consequential and causal events associated with motoneuron degeneration in amyotrophic lateral sclerosis (ALS). Current theories regarding the pathogenesis of motoneuron loss in ALS include, but are not limited to, central nervous system (CNS) excitotoxicity, autoimmune responses, mutations in the superoxide dismutase gene if familial ALS, na central or peripheral loss of trophic support. Interestingly, these events have also been shown to be involved in processes associated with programmed cell death, or apoptosis. The first Specific Aim of the present application is to study the expression of markers of programmed cell death in ALS autopsy material. This will be done by examining ALS and control spinal cord and motor cortex sections for cells exhibiting DNA fragmentation, a characteristic of apoptosis. In the second Specific Aim, we will use in situ hybridization and, in some cases, immunocytochemistry, to investigate the expression of certain cell death genes and their protein products in ALS tissue. The third Specific Aim will involve the screening of ALS cerebral spinal fluid (CSF) for the presence of immunoglobulins (e.g.Fas/Apo-1) known to be involved in programmed cell death. The outcome of these studies will determine the role of programmed cell death in ALS, and provide insight into potential therapeutic strategies to block this cell death process. One therapeutic approach currently being investigated is the treatment of ALS patients with neurotrophic factors. The rationale for using these molecules to treat ALS includes the understanding that neurotrophic factors promote neuronal survival and regrowth processes, and that they show some degree of selectivity for the populations of neurons in which they are effective. If trophic factor therapies are to be effective, it is essential to demonstrate that ALS motoneurons exhibit the capacity to respond to this treatment strategy. An initial step is the demonstration that ALS motoneurons express the receptors necessary for trophic factor signal transduction. The fourth Specific Aim will investigate the expression of the receptors for those trophic factors currently under investigation as therapeutic agents in the treatment of ALS. These receptors include the trks, which are the high-affinity receptors for the neurotrophins, and the alpha subunit of the CNTF receptor. We will use in situ hybridization to study the expression of specific mRNA encoding these receptors in ALS autopsy material. In addition, should adequate antibodies to the neurotrophic factor receptor proteins become available, we will incorporate the logical immunocytochemical experiments to compliment the study of mRNA expression. Overall, these studies are designed to advance our understanding of motoneuron degeneration in ALS, whether these motoneurons maintain the capacity to respond to certain trophic factor treatments, and demonstrate the potential role of programmed cell death in this devastating neurodegenerative disorder.