The Role of Calcyon in Synaptic Integration

Calcyon 在突触整合中的作用

基本信息

批准号：
6320696
负责人：
CLARE M BERGSON
金额：
$ 25.11万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-04-01 至 2005-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6320696
关键词：
autoradiography biological signal transduction calcium indicator calcium ion cell line confocal scanning microscopy dopamine receptor immunocytochemistry immunoprecipitation ion transport membrane proteins neural transmission phosphorylation protein protein interaction protein structure function receptor coupling synapses yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Obtaining a precise understanding of the mechanism(s) by which endogenous neuromodulators such as dopamine (DA) regulate glutamergic excitatory transmission could lead to better for patients afflicted with epilepsy, Parkinson's Disease, schizophrenia, and drug addiction. Activation of the D1 dopamine receptor subtype, in particular, modulates glutamate-induced neuronal excitability in frontal cortex, hippocampus and neostriatum. Little is known about molecular mechanism(s) by which D1 receptor signaling modulates excitatory transmission. We recently identified a single transmembrane protein designated Calcyon which interacts with D1 DA receptors and localizes to vesicles in dendritic spines. Via interaction with Calcyon, D1 receptors stimulate release of Ca++ from internal stores (Ca++i) in an activity-dependent manner in heterologous expression system. A similar activity-dependent, D1 receptor agonist stimulated response is detectable by Fura-2 Ca++ imaging of primary cultures of cortical and hippocampal neurons. Our overarching hypothesis is that Calcyon enhances D1 receptor signaling through Gq/11 by serving as a link connecting D1 receptors to IP3 regulated Ca++ stores. We will determine how D1 DA receptor signaling through Gq/11 is primed, and how Calcyon enhances D1 receptor-mediated IP3 signaling. Using Ca++ imaging and peptides to block the D1 receptor:Calcyon interaction, we will determine if Calcyon is required for D1 agonist stimulated Ca++i release in dendritic spines. We propose to identify other GPCRs and accessory proteins that interact with Calcyon by yeast two-hybrid and phage display screens. We will determine how these proteins are involved in regulating release of Ca++ from vesicular stores. The results of this research should provide insight into clinical strategies to selectively boost or dampen D1 receptor mediated neuromodulation of excitatory transmission.

描述（由申请人提供）：准确理解内源性神经调节剂如多巴胺 (DA) 的调节机制谷氨酸兴奋性传递可能会给患者带来更好的结果患有癫痫、帕金森病、精神分裂症和毒瘾。 D1 多巴胺受体亚型的激活尤其可以调节谷氨酸诱导额叶皮层、海马和新纹状体。关于 D1 受体的分子机制知之甚少信号传导调节兴奋性传递。我们最近确定了一个跨膜蛋白 Calcyon 与 D1 DA 受体相互作用并定位于树突棘的囊泡。通过与 Calcyon、D1 互动受体刺激内部储存 (Ca++i) 释放 Ca++ 异源表达系统中的活性依赖性方式。类似的活性依赖性 D1 受体激动剂刺激的反应可通过以下方式检测皮质和海马神经元原代培养物的 Fura-2 Ca++ 成像。我们的首要假设是 Calcyon 增强 D1 受体信号传导通过 Gq/11 作为连接 D1 受体与 IP3 调节的纽带 Ca++ 商店。我们将确定 D1 DA 受体信号如何通过 Gq/11 进行以及 Calcyon 如何增强 D1 受体介导的 IP3 信号传导。使用Ca++ 成像和肽来阻断 D1 受体：Calcyon 相互作用，我们将确定 D1 激动剂刺激 Ca++i 释放是否需要 Calcyon 树突棘。我们建议鉴定其他 GPCR 和辅助蛋白通过酵母双杂交和噬菌体显示屏与 Calcyon 相互作用。我们将确定这些蛋白质如何参与调节 Ca++ 的释放来自囊泡商店。这项研究的结果应该提供以下见解：选择性增强或抑制 D1 受体介导的临床策略兴奋性传递的神经调节。