RECOMBINANT GLUTAMATE RECEPTORS

为研究其结构和功能提出了实验方案。 重组谷氨酸受体的研究进展 爱马仕(alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate)和海人藻酸 子类型。天然受体和重组受体的功能特性 在非洲爪哇卵母细胞中表达的蛋白将通过电生理学进行比较 方法：研究方法。脑组织中AMPA或海人藻酸受体亚单位的水平 衍生的RNA将通过聚合酶链式反应(PCR)和 将在函数性质和可能的 子单元结构。这些相关性将通过测量 同源重组受体的功能特性研究 在脑部信使核糖核酸制剂中发现的这些亚基的数量。 多肽亚基特异性抗体的研究进展 在克隆的氨基酸序列中发现的氨基酸序列也被提出。有用 抗体将有助于直接探测天然的亚基结构 蛋白质印迹分析和高分辨率的受体 免疫定位。鉴定和鉴定激动剂和 在各个亚基的亚结构内的拮抗剂结合部位， 将有目的地改变亚基的结构 使用重组DNA技术。这些变化对 激动剂和拮抗剂的效力将用两个 非洲爪哇卵母细胞的电极电压钳。利用RAPID进行动力学研究 膜片钳瞬时阻断成纤维细胞的应用技术 表达的受体亚基将被用来剖析分子 配体结合和解离的机制。这些研究旨在 加深对神经元型AMPA和海人藻酸受体的了解 导致开发有用的治疗化合物，对 这些是感受器。AMPA和海人藻酸受体系统被牵连 在许多病理过程中，包括导致神经元损伤 由脑缺血、帕金森氏症、癫痫和精神病引起。

Experiments are proposed to study the structure and function of recombinant glutamate receptors of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) and kainate subtypes. The functional properties of native and recombinant receptors expressed in Xenopus oocytes will be compared using electrophysiological methods. The levels of AMPA or kainate receptor subunits in brain derived RNAs will be quantitated by polymerase chain reaction (PCR) and correlations will be made between functional properties and possible subunit structure. These correlations will be tested by measuring the functional properties of recombinant receptors expressed from identical amounts of the those subunits found in brain mRNA preparations. Development of subunit-specific antibodies directed against peptides found in the cloned amino acid sequences is also proposed. Useful antibodies will help directly probe the subunit structure of native receptors by Western blot analysis and high resolution immunolocalization. To identify and characterize the agonist and antagonist binding sites within the substructures of individual subunits, purposeful alterations in structure of the subunits will be introduced using recombinant DNA technology. The effects of these alterations on the potencies of agonists and antagonists will be measured with two electrode voltage clamp of Xenopus oocytes. Kinetic studies using rapid application techniques in patch clamped fibroblasts transiently expressing receptor subunits will be used to dissect the molecular mechanism of ligand binding and unbinding. These studies are designed to increase our understanding of neuronal AMPA and kainate receptors and lead toward the development of useful therapeutic compounds that act on these, receptors. The AMPA and kainate receptor systems are implicated in a number of pathological processes including neuronal damage resulting from cerebral ischemia, Parkinson's disease, epilepsy and psychosis.