Characterization of Human Trace Amine Receptors

人类痕量胺受体的表征

• 批准号: 7023844
• 负责人: KIM Arthur NEVE
• 金额: $ 14.76万
• 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 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7023844
• 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(申请人提供)：痕量胺是生物来源的非儿茶酚胺单胺，存在于脊椎动物神经系统中，浓度较低。一些微量的胺，如章鱼胺和酪胺，在无脊椎动物的神经系统中含量更丰富，它们作为神经递质的功能已经相当成熟。中枢神经系统中微量胺的异质性分布和快速周转，以及微量胺受体的存在，提示某些微量胺在哺乳动物中枢神经系统中也可能是神经递质或共递质。这一建议利用了最近对一个示踪胺受体(TAR)家族的DNA进行鉴定和分子克隆的优势。有趣的是，TAR聚集在人类6号染色体(6q23)的一个区域，许多研究表明该区域与精神分裂症有关。微量胺在神经精神障碍中的作用已被提出，TARS也是许多滥用药物的作用场所。这些发现提出了一种假设，即其中一种TAR的多态改变了蛋白质的功能，从而导致精神分裂症和/或药物滥用的症状。作为评估这一假说的第一步，我们提出了以下具体目标：1.在促进TAR在细胞表面表达的条件下，每种TAR将在哺乳动物细胞中稳定表达。将测试的三种策略是对受体进行遗传修饰，使用潜在的配体作为“药理伴侣”，以及抑制受体内化。2.TARs的激动剂和拮抗剂将通过测试潜在的单胺配体刺激或抑制循环AMP积累的能力以及激活稳定表达受体亚型的哺乳动物细胞中的丝裂原激活蛋白激酶的能力来确定。3.含有非同义SNPs的TAR变异体将在哺乳动物细胞中稳定表达，以确定该多态对细胞表面表达、配体结合和受体功能的影响。这些研究的理想结果将是确定所有编码全长受体的TAR在细胞表面表达的条件，确定每个TAR亚型的激动剂和拮抗剂，以及确定改变TAR亚型功能或配体结合的等位基因变体。