METHAMPHETAMINE EFFECTS VIA TRACE AMINE ASSOCIATED RECEPTOR 1

甲基苯丙胺通过微量胺相关受体 1 发挥作用

• 批准号: 7715483
• 负责人: GREGORY MICHAEL MILLER
• 金额: $ 1.95万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-05 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7715483
• 关键词: Affinity; Amines; Autoreceptors; Binding; Biological Assay; Brain; Brain region; Cells; Computer Retrieval of Information on Scientific Projects Database; Corpus striatum structure; Dopamine; Funding; Grant; H 89; In Vitro; Institution; Knockout Mice; Methamphetamine; Monkeys; Neurotransmitters; Norepinephrine; PKA inhibitor; Pathway interactions; Pharmaceutical Preparations; Phosphorylation; Research; Research Personnel; Resources; Role; Serotonin; Signal Transduction; Source; Synaptosomes; Thalamic structure; United States National Institutes of Health; Wild Type Mouse; brain cell; dopamine transporter; extracellular; inhibitor/antagonist; monoamine; noradrenaline transporter; psychostimulant; receptor; response; serotonin transporter; uptake

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The psychostimulant drug methamphetamine interferes with monoamine transporters to influence extracellular levels of monoamine neurotransmitters and alter brain monoaminergic tone. Trace amine-associated receptor 1 (TAAR1) is expressed in monoaminergic brain regions and is activated by methamphetamine in vitro. However, it is unclear whether TAAR1 is implicated in methamphetamine effects on monoamine transporters. This study used transfected HEK293 cells and brain synaptosomes to investigate the role of TAAR1 in methamphetamine modulation of the dopamine transporter (DAT), norepinephrine transporter (NET) and serotonin transporter (SERT). We found that TAAR1 activation by methamphetamine significantly inhibited uptake and promoted efflux of (3H)dopamine, (3H)norepinephrine and (3H)serotonin by DAT, NET and SERT, respectively, in transfected cells. Similarly, methamphetamine significantly inhibited uptake and induced efflux of (3H)dopamine and (3H)serotonin in striatal and (3H)norepinephrine in thalamic synaptosomes of monkeys and wild type mice, but it had no such effect in TAAR1 knockout mouse synaptosomes. Both the PKA inhibitor H89 and PKC inhibitor Ro32-0432 blocked uptake inhibition caused by methamphetamine, and Ro32-0432 blocked methamphetamine-induced efflux in wild type mouse synaptosomes. TAAR1 signaling in response to methamphetamine was unaffected by monoamine autoreceptors (D2s, alpha2A, alpha2B, 5HT1A and 5HT1B) co-expressed in transfected cells, and binding assays showed that methamphetamine had extremely low binding affinity for the monoamine autoreceptors in contrast to dopamine, norepinephrine and serotonin. Together, these results provide the first evidence that methamphetamine interacts with TAAR1 but not monoamine autoreceptors to alter uptake and efflux functions of brain monoamine transporters via phosphorylation-dependent pathways.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 精神刺激性药物甲基苯丙胺会干扰单胺转运蛋白，以影响单胺神经递质的细胞外水平并改变脑单胺能张力。痕量胺相关受体1（TAAR1）在单胺能脑区域表达，并在体外被甲基苯丙胺激活。但是，尚不清楚taAR1是否与甲基苯丙胺对单胺转运蛋白的作用有关。这项研究使用了转染的HEK293细胞和脑突触体来研究TAAR1在多巴胺转运蛋白（DAT），去甲肾上腺素转运蛋白（NET）和5-羟色胺转运蛋白转运蛋白（SERT）中的甲基苯丙胺调节中的作用。我们发现，通过甲基苯丙胺的TAAR1激活显着抑制了（3H）多巴胺，（3H）去甲肾上腺素的（3H）多巴胺和（3H）5-羟色胺的摄取，分别通过DAT，NET和SERT在转染的细胞中。同样，甲基苯丙胺在纹状体和野生型小鼠的丘脑突触体中显着抑制了（3H）多巴胺和（3H）5-羟色胺的摄取和（3H）5-羟色胺，但在TAAR1敲除小鼠突触中没有这种作用。 PKA抑制剂H89和PKC抑制剂RO32-0432都阻止了甲基苯丙胺引起的摄取抑制作用，RO32-0432阻断了野生型小鼠突触体中甲基苯丙胺诱导的外排。 TAAR1对甲基苯丙胺的信号传导不受单胺自​​感受器（D2S，Alpha2a，Alpha2a，Alpha2b，5HT1A和5HT1A和5HT1B）的影响，在转染的细胞中共表达，并且结合测定法显示，甲基苯胺对单氨基的结合性非常低的甲基氨基胺和含量的甲基氨基酶的相关性非常低和5-羟色胺。总之，这些结果提供了第一个证据，表明甲基苯丙胺与TAAR1相互作用，但没有单胺自身受体来改变脑单胺转运蛋白通过磷酸化依赖性途径的吸收和外排功能。