COCAINE EFFECTS ON NMDA RECEPTOR/PROTEIN INTERACTIONS

描述 这项提案的总体目标是检查推定的分子 与可卡因诱导的N-甲基-D- 谷氨酸受体的天冬氨酸（NMDA）亚型。可卡因给药 据报道可改变NMDA受体亚单位NR1的表达， 一种区域性的方式。最近的方法发展 提供检查可卡因对NMDA受体诱导作用的方法 具有增加的特异性的复合物，从而使得能够更全面地 评估蛋白质表达的变化。具体目标是 建议是：1）表征可卡因诱导的表达变化 以及NMDA受体亚基和剪接变体蛋白的定位-- 包括潜在的停药时间依赖性差异，2）进一步 探讨神经元型一氧化氮合酶（nNOS）、 钙调蛋白（CaM）和NMDA受体亚单位的推定机制 与可卡因诱导的NMDA受体表达变化相关， 3)评估与可卡因相关的NMDA受体功能的改变 通过nNOS活性测量的治疗。CaM、nNOS和NMDA的变化 受体亚基和剪接变体的表达将使用 免疫组织化学技术。免疫沉淀将用于 评价可卡因可能影响的CaM、nNOS和NMDA的破坏 受体相互作用可卡因对NMDA功能的影响 将使用nNOS活性测定来评估受体。数据的日期及时间为 这对于理解神经系统的分子和神经解剖学基础非常重要， 药物对神经传递和突触组织的影响， 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.