COCAINE EFFECTS ON NMDA RECEPTOR/PROTEIN INTERACTIONS

描述 这项提议的总体目标是研究假定的分子 可卡因诱导N-甲基-D-天冬氨酸的作用机制 谷氨酸受体的天冬氨酸(NMDA)亚型。可卡因的使用 据报道可以改变NMDA受体亚单位NR1的表达 一种特定于地区的方式。目前方法学的最新发展 提供检查可卡因对NMDA受体的影响的方法 复杂性增加了特异性，从而实现了更全面的 评估蛋白质表达的变化。这样做的具体目的是 建议是：1)表征可卡因诱导的表达变化 以及NMDA受体亚基和剪接变异体蛋白的定位-- 包括潜在的与撤资时间相关的差异，2)进一步 探讨神经元型一氧化氮合酶(NNOS)、 钙调素(CaM)和NMDA受体亚基可能的作用机制 与可卡因诱导的NMDA受体表达的变化有关，以及 3)评估与可卡因有关的NMDA受体功能的改变 治疗以nNOS活性为指标。CaM、nNOS和NMDA的变化 受体亚单位和剪接变异体的表达将用 免疫组织学技术。免疫沉淀法将用于 评估可卡因对CaM、nNOS和NMDA的影响 受体相互作用。可卡因对NMDA功能的影响 受体将使用nNOS活性测定进行评估。调查结果将是 对于理解脑脊髓炎的分子和神经解剖学基础很重要 药物对神经传递和突触组织的影响，并可能 此外，还有助于治疗的发展。

DESCRIPTION The overall goal of this proposal is to examine putative molecular mechanisms associated with cocaine-induced effects on the N-methyl-D- aspartate (NMDA) subtype of glutamate receptor. Administration of cocaine is reported to alter the expression of the NMDA receptor subunit, NR1, in a regionally specific manner. Recent methodological developments now provide the means to examine cocaine-induced effects on the NMDA receptor complex with increased specificity thus enabling a more comprehensive assess of changes in protein expression. The specific aims of this proposal are: 1) to characterize cocaine-induced changes in the expression and localization of NMDA receptor subunit and splice variant proteins-- including potential withdrawal time-dependent differences, 2) to further explore the relationship among neuronal nitric oxide synthase (nNOS), calmodulin (CaM), and NMDA receptor subunits for putative mechanisms associated with cocaine-induced changes in NMDA receptor expression, and 3) to assess alteration in NMDA receptor function associated with cocaine treatment as measured by nNOS activity. Changes in CaM, nNOS, and NMDA receptor subunit and splice variant expression will be investigated using immunohistological techniques. Immunoprecipitation will be used to evaluated possible cocaine-influenced disruption of CaM, nNOS and NMDA receptor interaction. Cocaine-induced changes in the function of NMDA receptors will be assessed using a nNOS activity assay. Findings will be important for understanding the molecular and neuroanatomical bases for drug effects on neurotransmission and synaptic organization, and may additionally contribute to therapeutic developments.