GLUTAMATE-GATED CHANNELS IN CENTRAL & PERIPHERAL NEURONS

中环谷氨酸门控通道

• 批准号: 2891848
• 负责人: James E Huettner
• 金额: $ 20.67万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2891848
• 关键词: calcium flux; dizocilpine; glutamate receptor; glutamates; hippocampus; kainate; laboratory rat; neural transmission; neurons; neuropharmacology; receptor expression; synapses; tetrodotoxin; tissue /cell culture; voltage /patch clamp

The long term objective of my work is to provide a better understanding of synaptic transmission by studying the operation of NMDA, AMPA, and kainate receptors, which form ion channels gated by the neurotransmitter glutamate. Another major goal is to uncover properties of these receptors that may allow for clinical intervention to prevent excitotoxic cell death. The experiments outlined in this proposal focus on the functional properties of kainate receptors. This glutamate receptor subtype is thought to regulate neuronal excitability and may contribute to excitotoxicity under certain conditions; but, currently much less is known about kainate receptors than about NMDA and AMPA receptors. Patch clamp techniques will be used to record the whole-cell add single channel currents evoked by excitatory amino acids in isolated neurons that express native glutamate receptors, and in heterologous cells that express cloned glutamate receptor subunits. Kainate receptors will be characterized with respect to their agonist and antagonist pharmacology, desensitization properties, current-voltage relations, relative permeability to calcium and monovalent ions, and susceptibility to modulation. For experiments on CNS neurons, NMDA and AMRA receptors will be blocked with the non- competitive antagonists MK-801 and GYKI 53655, respectively. A second major aim of this study is to test the hypothesis that kainate receptors regulate synaptic transmission. The action of kainate initially will be studied on synapses formed by hippocampal neurons grown in microcultures. In preliminary experiments, the agonists kainate and domoate have been shown to inhibit both excitatory and inhibitory synapses. The possible mechanisms that may underlie this inhibition will be examined in detail. We will seek to establish whether this action involves pre or postsynaptic kainate receptors and whether they are identical to the receptors that mediate kainate currents in these cells. In addition, we will seek to determine the conditions under which kainate receptors may be activated by the endogenous agonist, glutamate.

我工作的长期目标是更好地理解 通过研究NMDA，AMPA和Kainate的操作，突触传播 受体，形成由神经递质门控的离子通道 谷氨酸。另一个主要目标是发现这些受体的特性 这可能允许临床干预以防止兴奋性细胞 死亡。该提案中概述的实验重点是功能 海藻酸盐受体的特性。这种谷氨酸受体亚型是 被认为可以调节神经元兴奋性，并可能有助于 在某些条件下的兴奋性毒性；但是，目前知之甚少 关于海藻酸盐受体，大于NMDA和AMPA受体。补丁夹 技术将用于记录整个细胞添加单个通道 电流在表达的孤立神经元中引起的兴奋性氨基酸引起 天然谷氨酸受体和表达克隆的异源细胞 谷氨酸受体亚基。海藻酸盐受体将被表征 尊重其激动剂和拮抗剂药理学，脱敏 特性，电流关系，钙相对渗透性 和单价离子，以及对调节的敏感性。进行实验 CNS神经元，NMDA和AMRA受体将被非 - 竞争性拮抗剂MK-801和Gyki 53655。 这项研究的第二个主要目的是检验海藻酸盐的假设 受体调节突触传播。海藻酸盐的作用最初 将研究由生长的海马神经元形成的突触 微培养。在初步的实验中，激动剂海谷和 已显示抑制兴奋性和抑制性 突触。可能是这种抑制作用的可能机制将 详细检查。我们将寻求确定这项行动是否 涉及前或突触后海谷酸酯受体，以及它们是否是 与介导这些细胞中介导海谷电流的受体相同。 此外，我们将寻求确定毛主世界的条件 受体可以被内源性激动剂谷氨酸激活。