Modulation of Serotonergic Transporters

血清素转运蛋白的调节

• 批准号: 7574438
• 负责人: ANITA SIDHU
• 金额: $ 30.97万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-02 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7574438
• 关键词: Affinity; Anxiety Disorders; Binding; Binding Proteins; Biological Assay; Biology; Biotinylation; Brain; Brain region; Cell Culture Techniques; Cell membrane; Cell surface; Cells; Characteristics; Cocaine; Cognition Disorders; Complementary DNA; Complex; Cytoskeleton; Data; Development; Dopamine; Drug Addiction; Drug abuse; Drug usage; Event; Exposure to; Fluorescence Resonance Energy Transfer; Functional disorder; Genes; Grant; Homeostasis; Immunology; Immunoprecipitation; Inclusion Bodies; Knockout Mice; Laboratories; Lead; Lewy Bodies; Link; Mediating; Mental disorders; Microtubules; Molecular; Molecular Target; Mood Disorders; Moods; Mus; Mutation; Nature; Nerve; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Neurotransmitters; Nocodazole; Paclitaxel; Parkinson Disease; Pharmaceutical Preparations; Phosphorylation; Physiological; Process; Property; Protein Kinase C; Proteins; Rattus; Regulation; Relative (related person); Research Personnel; Role; Schizophrenia; Serotonin; Signal Transduction; Sleep; Synapses; Synuclein Family; Testing; Therapeutic; Vesicle; Wild Type Mouse; alpha synuclein; attenuation; depression; desensitization; dopamine transporter; ecstasy; genetic regulatory protein; heuristics; immunocytochemistry; indexing; inhibitor/antagonist; member; novel; novel therapeutics; presynaptic; programs; protein function; protein protein interaction; raphe nuclei; serotonin transporter; synaptic function; syntaxin 1A; trafficking; uptake

DESCRIPTION (provided by applicant): Serotonin [5-hydroxytryptamine, 5-HT] is a neurotransmitter, whose dysfunction is linked to the genesis of numerous psychiatric diseases, including mood disorders. The presynaptic serotonin transporters (SERTs) constitute the primary mechanism for re-uptake of 5-HT and termination of its signal in the synapse, and are high-affinity targets for addictive drugs including MDMA, cocaine and tricyclics, and SERT inhibitors constitute one of the largest groups of drugs used in the therapeutic treatment of depression. Little is known about the mode by which SERT function is regulated. Our preliminary data has identified a-synuclein, a presynaptic vesicle-associated protein of unknown function, as a modulator of SERT function, causing attenuation of normal transporter activity, indexed by diminished [3H]5HT uptake assays. The interaction between SERT and a-synuclein proceeds through the formation of stable protein:protein heteromeric complexes, and results in trafficking of SERT away from the cell surface of the plasma membrane. In this proposal we will study in detail the mechanisms which underline such regulation of transporter activity, using cells co-transfected with alpha-synuclein, and its subtypes, and the SERT cDNA, as well as in cultured primary neurons. In particular, the identity of the structural components which both facilitate and reverse a-synuclein effects on SERT function and trafficking, will be analyzed thoroughly, through a battery of studies to include, immunology, transporter assays, immunocytochemistry and FRET. We will also examine the participation of a-synuclein in both protein kinase C-mediated phosphorylation of SERT, and in transporter desensitization upon exposure to 5-HT. Our previous studies showing that a-synuclein can also modulate the function of the dopamine transporter, suggests that a primary function of this protein may be the regulation of monamine neurotransmitter homeostasis. Thus, our studies on SERT may have added heuristic and practical benefits for the understanding of other neurotransmitter transporters linked to mental illness and drug abuse. The identification of a-synuclein as a regulatory protein partner of SERT holds promise in the development of novel therapeutic strategies in the treatment of depression and drug addiction.

描述（由申请人提供）：5-羟色胺[5-羟色胺，5-HT]是一种神经递质，其功能障碍与包括情绪障碍在内的多种精神疾病的起源有关。突触前的5-羟色胺转运蛋白（SERTS）构成了重新摄取5-HT并在突触中终止其信号的主要机制，并且是成瘾性药物的高亲和力靶标，包括MDMA，可卡因和三环类药物，以及Sert抑制剂，以及用于抑郁症的最大药物。关于SERT函数的调节模式知之甚少。我们的初步数据已确定A-突触核蛋白，一种与未知功能的突触前囊泡相关的蛋白质，是SERT功能的调节剂，导致正常转运蛋白活性的衰减，并通过[3H] 5HT 5HT摄取分析索引。 SERT和A-突触核蛋白之间的相互作用是通过稳定蛋白质的形成：蛋白质异晶络合物的形成，并导致SERT从质膜的细胞表面运输。在此提案中，我们将使用与α-核蛋白及其亚型的细胞以及SERT cDNA以及培养的原发性神经元中共转染的细胞，详细研究这些机制，这些机制强调了转运蛋白活性的调节。特别是，将通过一系列研究彻底分析促进和反向A-突触核蛋白对SERT功能和运输的影响的结构成分的身份，包括免疫学，转运蛋白分析，Immunocycomyocytoryssess，Immunocytorystress和Fret。我们还将检查A-突触核蛋白在蛋白激酶C介导的SERT的磷酸化中，以及暴露于5-HT时的转运蛋白脱敏。我们先前的研究表明，A-突触核蛋白还可以调节多巴胺转运蛋白的功能，这表明该蛋白质的主要功能可能是孤原神经递质稳态的调节。因此，我们对SERT的研究可能为了解与精神疾病和药物滥用有关的其他神经递质转运蛋白的理解增加了启发式和实际的好处。鉴定A-核蛋白作为SERT的调节蛋白伴侣在抑郁症和药物成瘾治疗方面的新​​型治疗策略方面有希望。