Neurotransmitter Corelease

神经递质共释放剂

• 批准号: 9927697
• 负责人: ROBERT H EDWARDS
• 金额: $ 37.66万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9927697
• 关键词: Anatomy; Behavior; Behavioral; Cells; Corpus striatum structure; Coupled; Couples; Dopamine; Future; GIRK2 subunit, G protein-coupled inwardly-rectifying potassium channel; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Glutamate Receptor; Glutamate Transporter; Glutamates; Image; Measurement; Mental Depression; Midbrain structure; Molecular; Morphologic artifacts; Nervous system structure; Neurons; Neurotransmitters; Output; Physiological; Population; Property; Proteomics; Recycling; Role; Signal Transduction; Synaptic Vesicles; Vesicle; Work; dopaminergic neuron; in vivo; information processing; inward rectifier potassium channel; monoamine; pH gradient; postsynaptic; presynaptic; response; segregation; uptake; vesicular monoamine transporter

Accumulating evidence shows that many neurons release two classical neurotransmitters, but fundamental questions remain about the cellular basis for corelease, with important implications for its physiological role. In this proposal, we use the vesicular neurotransmitter transporters to elucidate the mechanisms involved in corelease. In previous work, we showed that glutamate corelease by midbrain dopamine neurons serves two distinct roles, one in vesicle filling with dopamine and the other as an independent signal. Although the effects on vesicle filling require colocalization of the vesicular monoamine transporter VMAT2 and vesicular glutamate transporter VGLUT2 on the same synaptic vesicles, anatomy has suggested some segregation as well, but with unclear physiological consequences. We now find that dopamine neurons release glutamate and dopamine with different properties. Release of the two transmitters differs in short-term depression and depends on different presynaptic Ca++ channels. Synaptic vesicles belong to pools that differ in response to stimulation but these differences have been attributed to extrinsic factors such as cytoskeletal association. We now show that they also differ in composition because they contain different transmitters and release them with different properties. Through this mechanism, a single neuron can deconvolve its input into two distinct outputs. The long-term objectives of this project are to elucidate the cellular and molecular basis for neurotransmitter corelease and determine its role in information processing. The strategy is to use the vesicular transporters to characterize the different vesicle populations. Specifically, we will 1) compare monoamine and glutamate release by imaging VMAT2 and VGLUT2 in live neurons; 2) determine how VMAT2 and VGLUT2 target to distinct vesicle populations; 3) characterize the composition of monoamine and glutamate SVs by proteomics. The results will provide basic information about the organization of neurons, with direct relevance for corelease by other cells, but also for release by all neurons.

积累的证据表明，许多神经元释放了两个经典的神经递质，但 关于Corelease的细胞基础，基本问题仍然具有重要意义 生理角色。在此提案中，我们使用囊泡神经递质转运蛋白来阐明 涉及的机制。在以前的工作中，我们证明了中脑的谷氨酸corelease 多巴胺神经元具有两个不同的角色，一个在囊泡中充满多巴胺，另一个是独立的 信号。尽管对囊泡填充的影响需要囊泡单胺转运蛋白共定位 VMAT2和囊泡谷氨酸转运蛋白vglut2在同一突触囊泡上，解剖学表明 有些种族隔离，但有不清楚的生理后果。 现在，我们发现多巴胺神经元释放具有不同特性的谷氨酸和多巴胺。发布 在短期抑郁症中，两个发射器中有不同，取决于不同的突触前CA ++通道。 突触囊泡属于池的池，但这些差异已归因于 对于外部因素，例如细胞骨架关联。我们现在表明它们在构图上也有所不同，因为 它们包含不同的发射器，并以不同的特性释放它们。通过这种机制，一个 神经元可以将其输入分解为两个不同的输出。该项目的长期目标是阐明 神经递质取流的细胞和分子基础，并确定其在信息处理中的作用。 该策略是使用囊泡转运蛋白来表征不同的囊泡群体。具体来说， 我们将 1）通过成像VMAT2和VGLUT2在活神经元中比较单胺和谷氨酸释放； 2）确定VMAT2和VGLUT2如何靶向不同的囊泡群体； 3）通过蛋白质组学表征单胺和谷氨酸SV的组成。 结果将提供有关神经元组织的基本信息，并与Corelease直接相关 通过其他细胞，但也用于所有神经元的释放。