SCREENING ASSAY FOR EXCITATORY AMINO ACID ANTAGONISTS

兴奋性氨基酸(EaAs)与糖尿病的发病机制有关。 包括癫痫在内的许多人类神经疾病， 缺血性脑损伤和神经退行性疾病，如 亨廷顿病和阿尔茨海默病。EaaS在以下方面的作用 这些疾病似乎涉及异常增强的激活。 导致兴奋性毒性细胞损伤的EAA受体 神经元死亡。最近来自动物模型的证据表明 这些受体的选择性拮抗剂可以有效地 受体过度刺激和后续细胞的预防 死亡。这表明，EAA拮抗剂可以穿透 血脑屏障并选择性地作用于一个或多个EAA 受体亚型可能在临床上具有重要意义。 治疗涉及EAA神经毒性机制的疾病。 目前，EAA受体的初步鉴定 抗激动剂需要耗时且复杂的分析 使用活的动物或动物组织。我们建议开发一种 简化的检测系统，灵敏、可靠、快速，以及 这不需要持续的动物祭祀。在第I阶段， 在这个项目中，我们将鉴定和描述神经细胞系 表达一组新型神经细胞的功能性EAA受体 以前没有表现出这些特征的细胞系 感受器。这些品系是通过引进 利用逆转录病毒载体将癌基因导入原代脑组织。在……里面 第二阶段，我们将使用一种筛选方法，使用的细胞系包括 EAA受体用于从天然药物中鉴定新的EAA拮抗剂 来源(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.