MOLECULAR BASIS OF OPIOID PHARMACODYNAMICS

阿片类药物药效学的分子基础

• 批准号: 2116118
• 负责人: Charles E Inturrisi
• 金额: $ 10.69万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2116118
• 关键词: PC12 cells; adrenal medulla; analgesia; central nervous system; drug addiction; drug addiction antagonist; drug tolerance; endogenous opioid; excitatory aminoacid; gene induction /repression; genetic regulatory element; genetically modified animals; glucocorticoids; hamsters; human genetic material tag; laboratory mouse; laboratory rat; neurotransmitter receptor; neurotransmitters; nitric oxide synthase; nucleic acid sequence; opioid receptor; pharmacogenetics; pharmacokinetics; protooncogene; tissue /cell culture

This is a request for an ADAMHA Research Scientist Award. The application requests funds to allow me to pursue the acquisition of techniques of molecular biology and neuroscience and their application to 2 projects. The first project will examine by solution hybridization, Northern blot analysis, in situ hybridization and immunocytochemical techniques the mRNA and peptide levels for proenkephalin (Penk) prodynorphin NMDA receptor, nitric oxide synthase, c-fos and tyrosine hydroxylase in 15 select areas that are associated with opioid analgesia, tolerance and dependence. These studies will be conducted in animals made tolerant and/or dependent with mu, kappa and delta opioids and in the presence and absence of drugs (receptor antagonists and nitric oxide synthase inhibitors) which attenuate or prevent the development of opioid tolerance and dependence. In addition, a subtractive hybridization approach will be used to clone and identify proteins altered by opioid tolerance and dependence. The second project will define the factors including membrane depolarizing,second messenger systems and glucocorticoid that regulate the hamster, rat and human Penk genes. Glucocorticoid response elements in the hamster, rat and human Penk genes will be identified by cloning selected DNA fragments. These projects require the acquisition and application of a number of molecular and neuroscience techniques including in situ hybridization, subtractive hybridization, tissue culture, fusion gene construction, gene transfer, gel-shift mobility assays and transgenic analysis. The acquisition of these techniques will result from collaborations with scientists with expertise in each of these areas who have agreed to support the acquisition and transfer of these techniques to my laboratory. This RSA award will allow me the opportunity to reduce my teaching and administrative responsibilities and increase the time I can devote to enhancing my scientific skills.

这是 ADAMHA 研究科学家奖的请求。应用 请求资金让我能够获取以下技术 分子生物学和神经科学及其在 2 个项目中的应用。 第一个项目将通过溶液杂交、Northern blot 进行检查 分析、原位杂交和免疫细胞化学技术 和脑啡肽原 (Penk) 强啡肽原 NMDA 受体的肽水平， 15 个选定区域的一氧化氮合酶、c-fos 和酪氨酸羟化酶 与阿片类药物的镇痛、耐受性和依赖性相关。这些 研究将在耐受和/或依赖的动物中进行 mu、kappa 和 delta 阿片类药物以及存在或不存在药物的情况 （受体拮抗剂和一氧化氮合酶抑制剂） 减轻或防止阿片类药物耐受和依赖性的发展。 此外，将使用消减杂交方法来克隆 并鉴定因阿片类药物耐受性和依赖而改变的蛋白质。 第二个项目将定义包括膜在内的因素 去极化、第二信使系统和糖皮质激素调节 仓鼠、大鼠和人类 Penk 基因。糖皮质激素反应元件 仓鼠、大鼠和人类 Penk 基因将通过克隆选定进行鉴定 DNA 片段。 这些项目需要获取和应用许多 分子和神经科学技术，包括原位杂交， 消减杂交、组织培养、融合基因构建、基因 转移、凝胶迁移迁移率测定和转基因分析。这 这些技术的获得将通过与 在这些领域具有专业知识的科学家们同意 支持获取这些技术并将其转移到我的实验室。 这个 RSA 奖项将使我有机会减少我的教学和 行政职责并增加我可以投入的时间 提高我的科学技能。