SCREENING ASSAY FOR EXCITATORY AMINO ACID ANTAGONISTS

兴奋性氨基酸拮抗剂的筛选测定

• 批准号: 3504298
• 负责人: JAMES B FISCHER
• 金额: $ 4.99万
• 依托单位: CAMBRIDGE NEUROSCIENCE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1989-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3504298
• 关键词: antibody receptor; binding proteins; bioassay; biotechnology; calcium transporting ATPase; cell bank /registry; glutamates; kynurenate; membrane activity; membrane channels; messenger RNA; neural information processing; neural transmission; neurons; neuropharmacologic agent; neurotoxins; receptor expression

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.

兴奋性氨基酸 (EAA) 与以下疾病的病理学有关： 许多人类神经系统疾病，包括癫痫， 缺血性脑损伤和神经退行性疾病，例如 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 导致兴奋性毒性细胞损伤的 EAA 受体 神经元死亡。 来自动物模型的最新证据表明 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的细胞 死亡。 这表明 EAA 拮抗剂可穿透 血脑屏障并选择性地作用于一个或多个 EAA 受体亚型可能具有重要的临床意义 治疗涉及 EAA 神经毒性机制的疾病。 目前，初步鉴定EAA受体 拮抗剂需要耗时且复杂的测定 使用活体动物或动物组织。 我们建议开发一个 简化的检测系统灵敏、可靠、快速，并且 这不需要持续的动物牺牲。 在第一阶段 在这个项目中，我们将识别和表征神经元细胞系 表达来自一组新的神经细胞的功能性 EAA 受体 以前没有针对这些特征进行过表征的细胞系 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，我们将使用含有细胞系的筛选 EAA 受体可从天然产物中鉴定新型 EAA 拮抗剂 来源（例如蜘蛛毒液）和合成化合物 CNS 研究中心的分析。