AMPA DESENSITIZATION AND HYPOXIC/ISCHEMIC NEURONAL INJURY

AMPA 脱敏和缺氧/缺血性神经元损伤

• 批准号: 6243802
• 负责人: KELVIN A YAMADA
• 金额: $ 12.96万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-01 至 1998-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6243802
• 关键词: cerebral ischemia /hypoxia; cytotoxicity; glutamate receptor; kainate; laboratory rat; neuroprotectants; protein structure function; quinazolines; receptor binding; receptor expression; receptor sensitivity; thiazide; tissue /cell culture; voltage /patch clamp

Glutamatergic neurotransmitter systems are widely distributed in the mammalian CNS, and they are involved in excitatory synaptic transmission, synaptic plasticity, and in neural development. Glutamate receptors are also involved in neural injury from hypoxia-ischemia (stroke). This project will concentrate upon the AMPA subset of glutamate receptors and its role in hypoxic-ischemic neuronal injury. AMPA receptors mediate rapid, excitatory signalling between central neurons, and exhibit the property of rapid desensitization. This property of AMPA receptors is studied with patch-clamp electrophysiology in conjunction with pharmacological manipulation of AMPA desensitization, which has advanced significantly following the recent discovery in this laboratory of several drugs that specifically alter the desensitization properties of AMPA receptors. Concommitantly, AMPA receptors have emerged as important mediators of neuronal injury during experimental ischemia, producing great interest in understanding the modulatory factors (such as desensitization) for the purpose of targeting this receptor for therapeutic intervention in the treatment of stroke in an analogous fashion applied to the different sites on the NMDA receptor for the treatment of stroke and epilepsy or the GABA receptor for the treatment of epilepsy and anxiety disorders. The objectives of this proposal are to use patch-clamp electrophysiology in conjunction with receptor pharmacology and molecular biology in cell culture systems, cell lines expressing glutamate receptors, explant cultures of brain slices, and acutely prepared brain slices to identify regulatory mechanisms of AMPA receptor desensitization, and to determine how desensitization influences neuronal vulnerability to hypoxic-ischemic damage. Validation of the influence of these mechanisms upon hypoxic-ischemic neuronal damage will be examined in cultured cortical and hippocampal neurons, and hippocampal explant cultures subjected to oxygen-glucose deprivation ('in vitro ischemia'), as well as in rodents subjected to focal and global ischemia in order to expand these principles to more intact systems and live experimental animals. The long-term goal of this proposal is to enhance our understanding of the cellular mechanisms underlying the pathophysiology of stroke, and to expand and improve available means to treat patients who suffer from strokes.

谷氨酸能神经递质系统广泛分布于 哺乳动物中枢神经系统，它们参与兴奋性突触传递， 突触可塑性和神经发育。 谷氨酸受体是 还涉及缺氧缺血（中风）引起的神经损伤。 这 该项目将集中于谷氨酸受体的 AMPA 子集和 其在缺氧缺血性神经元损伤中的作用。 AMPA 受体介导 中枢神经元之间的快速、兴奋性信号传导，并表现出 具有快速脱敏的特性。 AMPA 受体的这一特性是 结合膜片钳电生理学进行研究 AMPA 脱敏的药理学操作，已取得进展 显着遵循该实验室最近的发现 几种专门改变脱敏特性的药物 AMPA 受体。 与此同时，AMPA 受体也变得越来越重要。 实验性缺血期间神经元损伤的介质，产生 对了解调节因素（例如 脱敏）以靶向该受体 中风治疗中的治疗干预类似 时尚应用于 NMDA 受体的不同位点 中风和癫痫的治疗或GABA受体的治疗 癫痫和焦虑症。 该提案的目标是 将膜片钳电生理学与受体结合使用 细胞培养系统、细胞系中的药理学和分子生物学 表达谷氨酸受体、脑切片的外植体培养物，以及 精心准备的脑切片以确定 AMPA 的调节机制 受体脱敏，并确定脱敏如何影响 神经元易受缺氧缺血损伤。 验证 这些机制对缺氧缺血性神经元损伤的影响 在培养的皮质和海马神经元中进行检查，并在海马 外植体培养物经受氧-葡萄糖剥夺（“体外 缺血'），以及遭受局灶性和全身性缺血的啮齿类动物 为了将这些原则扩展到更完整的系统和生活 实验动物。 该提案的长期目标是加强 我们对细胞机制的理解 中风的病理生理学，并扩大和改进可用的手段 治疗中风患者。