Trace Amine Receptors in Non Human Primates

非人类灵长类动物中的痕量胺受体

• 批准号: 7049449
• 负责人: GREGORY MICHAEL MILLER
• 金额: $ 28.88万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7049449
• 关键词: Macaca mulatta; amines; amphetamines; animal tissue; antiantibody; biological signal transduction; cell surface receptors; cocaine; immunocytochemistry; in situ hybridization; messenger RNA; molecular cloning; neuropharmacology; polymerase chain reaction; protein quantitation /detection; protein structure function; receptor binding; receptor expression

DESCRIPTION (provided by applicant): Trace amines, distinguishable from the well-characterized biogenic amines, have been known to exist in mammalian brain for over 25 years. The recent discovery of a family of mammalian trace amine receptors (TARs) presents a significant new resource for exploring the physiological and pharmacological relevance of trace amines in brain (Borowsky et al, 2001). In conjunction with this finding is the unanticipated discovery that drugs of abuse, ranging from amphetamine, MDMA, LSD, as well as dopamine stimulate rat TAR1-mediated cAMP formation at pharmacologically relevant concentrations (Bunzow et al, 2001). Accordingly, trace amine receptors may directly or indirectly contribute to psychostimulant and hallucinogenic drug effects. As human and rodent trace amine receptors diverge in structure, subtype number and brain distribution, we postulate that non-human primates will provide a more suitable model for uncovering the physiological and pharmacological relevance of trace amine subtypes. In this regard, pilot studies indicate a 96% sequence identity for TAR1 in non-human primate and human. To establish fundamental information needed to explore TAR subtype function, we will investigate non-human primate TAR subtype structure, pharmacology, signal transduction and brain distribution. Aim 1 will determine whether human TAR subtypes (TAR1, 3, 4, 5) and structurally related orphan receptors (GPR57, GPR58 and PNR) exist in nonhuman primates. Based on these results, Aim 2 will utilize luciferase assays sensitive to different signal transduction pathways to uncover agonists as well as antagonists that block agonist-induced receptor activation. To characterize the pharmacological profile of these receptors, radioreceptor assays will be developed from lead compound(s) identified from luciferase assays. Aim 3 will map TAR subtype mRNA and protein distribution in primate brain using in situ hybridization, real-time RT-PCR and immunohistochemistry, to discern which receptor subtypes are expressed in brain for further exploration. Our pilot studies suggest that primate TARs are direct targets of psychostimulant drugs of abuse in brain. The proposed studies will form the basis for investigating the physiological and pharmacological relevance of trace amine receptors in a primate model, and may provide novel leads for developing therapeutic agents to treat addiction and possibly neuropsychiatric disorders.

描述（由申请人提供）： 痕量胺与已充分表征的生物胺不同，已知在哺乳动物大脑中存在超过 25 年。最近发现的哺乳动物微量胺受体（TAR）家族为探索大脑中微量胺的生理和药理学相关性提供了重要的新资源（Borowsky 等，2001）。与这一发现相结合的是一个意想不到的发现，滥用药物，包括安非他明、MDMA、LSD 以及多巴胺，会在药理学相关浓度下刺激大鼠 TAR1 介导的 cAMP 形成（Bunzow 等，2001）。因此，微量胺受体可能直接或间接地促进精神兴奋和致幻药物作用。由于人类和啮齿动物的微量胺受体在结构、亚型数量和大脑分布方面存在差异，我们假设非人类灵长类动物将为揭示微量胺亚型的生理和药理学相关性提供更合适的模型。在这方面，初步研究表明 TAR1 在非人类灵长类动物和人类中具有 96% 的序列同一性。为了建立探索 TAR 亚型功能所需的基本信息，我们将研究非人类灵长类动物 TAR 亚型结构、药理学、信号转导和大脑分布。目标 1 将确定非人类灵长类动物中是否存在人类 TAR 亚型（TAR1、3、4、5）和结构相关的孤儿受体（GPR57、GPR58 和 PNR）。基于这些结果，Aim 2将利用对不同信号转导途径敏感的荧光素酶测定来发现激动剂以及阻断激动剂诱导的受体激活的拮抗剂。为了表征这些受体的药理学特征，将从荧光素酶测定中鉴定出的先导化合物开发放射受体测定。目标 3 将利用原位杂交、实时 RT-PCR 和免疫组织化学绘制灵长类动物大脑中 TAR 亚型 mRNA 和蛋白质的分布图，以辨别哪些受体亚型在大脑中表达，以供进一步探索。我们的初步研究表明，灵长类 TAR 是大脑中精神兴奋药物滥用的直接目标。拟议的研究将为研究灵长类动物模型中痕量胺受体的生理和药理学相关性奠定基础，并可能为开发治疗成瘾和可能的神经精神疾病的治疗药物提供新的线索。