Characterization of Human Trace Amine Receptors

人类痕量胺受体的表征

• 批准号: 6923377
• 负责人: KIM Arthur NEVE
• 金额: $ 12.6万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6923377
• 关键词: adenosine monophosphate; amines; cell surface receptors; immunocytochemistry; mitogen activated protein kinase; molecular cloning; octopamine; protein structure function; receptor binding; receptor expression; single nucleotide polymorphism; tissue /cell culture; transfection /expression vector; western blottings

DESCRIPTION (provided by applicant): Trace amines are biogenic non-catecholamine monoamines that are present at low concentrations in the vertebrate nervous system. Some trace amines, such as octopamine and tyramine, are more abundant in invertebrate nervous systems, where their function as neurotransmitters is fairly well established. The heterogeneous CNS distribution and rapid turnover of trace amines, together with the existence of trace amine receptors, suggest that some trace amines may also be neurotransmitters or co-transmitters in the mammalian CNS. This proposal takes advantage of the recent identification and molecular cloning of the DNA for a family of trace amine receptors (TARs). Interestingly, the TARs are clustered in a region of human chromosome 6 (6q23) where linkage to schizophrenia has been suggested by a number of studies. A role for trace amines in neuropsychiatric disorders has been proposed, and the TARS are also sites of action for many abused drugs. These findings suggest the hypothesis that a polymorphism in one of the TARs changes the function of the protein and thus contributes to the symptoms of schizophrenia and/or to drug abuse. As an initial step towards evaluating this hypothesis we propose the following specific aims: 1. Each TAR will be stably expressed in mammalian cells under conditions that promote its expression on the cell surface. Three strategies to be tested are genetic modification of the receptors, the use of potential ligands as "pharmacological chaperones", and inhibition of receptor internalization. 2. Agonists and antagonists for the TARs will be identified by testing the ability of potential monoamine ligands to stimulate or inhibit cyclic AMP accumulation and to activate mitogen-activated protein kinases in mammalian cells stably expressing the receptor subtypes. 3. TAR variants with non-synonymous SNPs will be stably expressed in mammalian cells to determine the effect of the polymorphism on cell surface expression, ligand binding, and function of the receptor. The ideal outcome of these studies would be to identify conditions under which all TARs encoding full-length receptors are expressed on the cell surface, to identify agonists and antagonists for each TAR subtype, and to identify allelic variants that alter the function or ligand binding of a TAR subtype

描述（由申请人提供）：痕量胺是生物源性非儿茶酚胺单胺，在脊椎动物神经系统中以低浓度存在。一些微量胺，如章鱼胺和酪胺，在无脊椎动物的神经系统中更丰富，它们作为神经递质的功能相当完善。微量胺在中枢神经系统的分布不均匀、代谢迅速以及微量胺受体的存在，提示某些微量胺可能也是哺乳动物中枢神经系统的神经递质或共递质。该提案利用了最近对痕量胺受体（TARs）家族DNA的鉴定和分子克隆。有趣的是，TARs聚集在人类6号染色体（6 q23）的一个区域，许多研究表明该区域与精神分裂症有关。微量胺在神经精神疾病中的作用已经被提出，并且TARS也是许多滥用药物的作用部位。这些研究结果表明，一个多态性在一个焦油改变蛋白质的功能，从而有助于精神分裂症的症状和/或药物滥用的假设。作为评估这一假设的第一步，我们提出了以下具体目标：1。每种TAR将在促进其在细胞表面上表达的条件下在哺乳动物细胞中稳定表达。三个策略进行测试的受体的遗传修饰，使用潜在的配体作为“药理学伴侣”，抑制受体内化。2.通过测试潜在的单胺配体刺激或抑制环AMP积累和激活稳定表达受体亚型的哺乳动物细胞中的促分裂原活化蛋白激酶的能力，鉴定TAR的激动剂和拮抗剂。3.具有非同义SNP的TAR变体将在哺乳动物细胞中稳定表达，以确定多态性对细胞表面表达、配体结合和受体功能的影响。这些研究的理想结果将是鉴定编码全长受体的所有TAR在细胞表面上表达的条件，鉴定每种TAR亚型的激动剂和拮抗剂，并鉴定改变TAR亚型的功能或配体结合的等位基因变体