Development of Fluorescent False Neurotransmitters

荧光假神经递质的开发

基本信息

批准号：
10636618
负责人：
DALIBOR SAMES
金额：
$ 61万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-18 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10636618
关键词：
Acute Address Adrenergic Receptor Amphetamines Animals Anxiety Attention Attention deficit hyperactivity disorder Axon Behavior Behavioral Biological Assay Brain Catecholamines Cell Culture Techniques Cells Central Nervous System Cerebellum Chemistry Cognition Complement Complex Corpus striatum structure Development Diffusion Dopamine Dopamine Receptor Drug Addiction Drug Modulation Environment Excitatory Synapse Fiber Fluorescence Fright Glutamates Hippocampus Image Imaging Device Individual Inhibitory Synapse Interdisciplinary Study Ion Channel Kinetics Label Learning Locomotion Measures Membrane Memory Mental Depression Methods Molecular Molecular Probes Monitor Mus Nervous System Neurobiology Neuropharmacology Neurosciences Neurotransmitters Norepinephrine Optical reporter Optics Parkinson Disease Pharmaceutical Preparations Photometry Photons Play Post-Traumatic Stress Disorders Process Property Proteins Psychotropic Drugs Quinolones Reporter Reporting Research Support Rewards Role Safety Schizophrenia Sensory Series Serotonin Signal Transduction Site Slice Stimulus Synapses Synaptic Vesicles Synaptic plasticity System Tail Tracer Varicosity Viral brain tissue combinatorial design ecstasy extracellular imaging capabilities imaging modality imaging probe in vivo insight microscopic imaging mind control monoamine multiphoton imaging multiphoton microscopy nervous system disorder neuropsychiatric disorder neurotransmission noradrenergic novel imaging technique optogenetics pharmacologic postsynaptic presynaptic programs psychostimulant receptor response sensor serotonin transporter small molecule source localization substance use temporal measurement tool transmission process uptake vesicular monoamine transporter 2

项目摘要

SUMMARY Development of Fluorescent False Neurotransmitters Monoamine neurotransmission plays important roles in modulation of excitatory and inhibitory synapses in the central nervous system and thus provides essential fine tuning of behavior in response to a changing environment. Consequently, aberrations in monoamine neurotransmission underlie many neurological and neuropsychiatric disorders including Parkinson’s disease, schizophrenia, ADHD, depression, and drug addiction. We have established an interdisciplinary research program focused on development of new molecular probes to study monoamine neurotransmission on a synaptic level. Specifically, we have introduced a conceptually new class of probes, termed “fluorescent false neurotransmitters” (FFNs), that act as optical tracers of dopamine and norepinephrine. FFNs provide the first experimental means to image synaptic vesicle content uptake and release at individual presynaptic varicosities in brain tissue and in living animals. Using FFNs and multiphoton microscopy imaging we have made several unexpected findings, including the discovery of silent dopaminergic and noradrenergic synapses, activity-dependent super-charging of synaptic vesicles, and mechanistic insights on psychostimulant drugs. In this renewal application, we propose to expand the scope of FFNs to serotonin synapses, the third major monoamine system, on the basis of proof-of-concept studies. We will also develop new imaging and photon-counting methods based on the combinatorial use of FFNs as presynaptic probes with emerging genetically encoded monoamine reporters as postsynaptic sensors to provide multi-parameter synaptic readouts in living mice. In addition to complementing each FFN class with the corresponding sensor (for example, dopamine FFN and dopamine sensor), mixing and matching FFNs and sensors each for a different monoamine will enable simultaneous examination of two neurotransmitter systems. This in turn will enable the study of complex processes such as co-transmission and neurotransmitter switching on a synaptic level. We will also examine the effect of monoamine releasing agents on synaptic neurotransmitter dynamics at the dopamine, norepinephrine, and serotonin axonal release sites to probe both the vesicular pools in active and silent varicosities and the mechanisms of action of these psychoactive drugs. This application addresses a major unmet need in developing molecular tools and methods for imaging multiple neurotransmitter systems in living animals with adequate spatial and temporal resolution.

概括荧光假神经递质的发展单胺神经传递在调节兴奋和抑制突触中起着重要作用中枢神经系统，因此为响应变化提供了基本的行为调整环境。因此，单胺神经传递的畸变是许多神经系统和神经精神疾病，包括帕金森氏病，精神分裂症，多动症，抑郁症和药物瘾。我们已经建立了一个跨学科研究计划，旨在发展新的分子问题在合成水平上研究单胺神经传递。具体来说，我们介绍了从概念上讲，一种新的问题，称为“荧光假神经递质”（FFNS），它是光学的多巴胺和去甲肾上腺素的示踪剂。 FFN提供了图像突触囊泡的第一种实验手段在脑组织和活体动物中的单个突触前静脉曲张中的含量吸收和释放。使用 FFN和多光子显微镜成像，我们做出了几个意外的发现，包括发现沉默多巴胺能和去甲肾上腺素能的突触，依赖于突触蔬菜的超级充电，以及关于精神刺激药物的机械见解。在此续订应用程序中，我们建议 FFN与5-羟色胺突触的范围，第三个主要的单胺系统，基于概念证明研究。我们还将根据组合使用的组合使用 FFN作为突触前问题，普遍编码的单胺记者作为突触后传感器在活小鼠中提供多参数突触读数。除了完成每个FFN课程外相应的传感器（例如多巴胺FFN和多巴胺传感器），混合和匹配的FFN和每个传感器的单体胺都可以同时检查两个神经递质系统。这反过打开突触水平。我们还将检查单胺释放剂对突触的影响多巴胺，去甲肾上腺素和5-羟色胺轴突释放位点的神经递质动力学均可探测活跃和沉默的静脉曲张中的囊泡池以及这些精神活性药物的作用机制。该应用程序解决了开发成像的分子工具和方法的主要未满足需求具有足够空间和临时分辨率的活动物中的多个神经递质系统。