SCREENING ASSAY FOR EXCITATORY AMINO ACID ANTAGONISTS

兴奋性氨基酸（EAA）参与了 许多人类神经系统疾病，包括癫痫， 缺血性脑损伤和神经变性疾病， 亨廷顿舞蹈病和阿尔茨海默病。 EAA的作用 这些疾病似乎涉及异常增强的激活 EAA受体，导致兴奋性细胞损伤， 神经元死亡 最近的动物模型证据表明 这些受体的选择性拮抗剂可以有效地 防止受体过度刺激和随后细胞 死亡 这表明，EAA拮抗剂，穿透 血脑屏障，并选择性地作用于一个或多个EAA 受体亚型可能是主要的临床重要性， 治疗涉及EAA神经毒性机制的疾病。 目前，EAA受体的初步鉴定 拮抗剂需要耗时且复杂的测定 使用活的动物或动物组织。 我们建议发展一个 灵敏、可靠和快速的简化测定系统，以及 这不需要持续的动物牺牲。 一阶段 本项目将鉴定和表征神经元细胞系 从一组新的神经细胞中表达功能性EAA受体， 细胞系，以前没有被表征为这些 受体。 这些线路是通过引入 使用逆转录病毒载体将癌基因导入原代脑组织。 在 第二阶段，我们将使用含有以下物质的细胞系进行筛选： EAA受体以从天然EAA受体中鉴定新型EAA拮抗剂 源（例如，蜘蛛毒液）和合成化合物 CNS Research的分析。

Excitatory amino acids (EAAs) are implicated in the pathology of a number of human neurological disorders, including epilepsy, ischemic brain damage, and neurodegenerative disases such as Huntington's disease and Alzheimer's disease. The role of EAAs in these disorders appears to involve abnormally enhanced activation of EAA receptors that leads to excitotoxic cell damage and neuronal death. Recent evidence from animal models indicates that selective antagonists for these receptors can be effective in the prevention of receptor over-stimulation and subsequent cell death. This suggests that EAA antagonists that penetrate the blood-brain barrier and act selectively at one or more of the EAA receptor subtypes may be of major clinical importance for the treatment of disorders involving EAA neurotoxic mechanisms. Currently, preliminary identification of EAA receptor anatagonists requires time-consuming and complicated assays using live animals or animal tissue. We propose to develop a simplified assay system that is sensitive, reliable, and rapid, and which does not require continual animal sacrifice. In Phase I of this project we will identify and characterize neuronal cell lines expressing functional EAA receptors from a novel group of neural cell lines that have not previously been characterized for these receptors. These lines were established by the introduction of oncogenes into primary brain tissues using retroviral vectors. In Phase II, we will use a screens employing cell lines containing EAA receptors to identify novel EAA antagonists from natural sources (e.g., spider venoms) and synthetic compounds under analysis at CNS Research.