Spectra-Physics Mai Tai HP Laser

Spectra-Physics 迈泰 HP 激光器

• 批准号: 7794473
• 负责人: David Sulzer
• 金额: $ 19.58万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-25 至 2011-03-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794473
• 关键词: Acute; Attention deficit hyperactivity disorder; Brain; Chemistry; Collaborations; Data; Decision Making; Dopamine; Dopamine D2 Receptor; Dopamine Receptor; Drug abuse; Energy Transfer; Event; Fluorescence; Funding; Generations; Habits; Image; Individual; Interdisciplinary Study; Joints; Kinetics; Laboratories; Lasers; Life; Light; Measures; Molecular; Neurons; Neurosciences; Neurotransmitters; Optical Methods; Optics; Parkinson Disease; Pharmacology; Physics; Presynaptic Terminals; Research; Resolution; Schizophrenia; Side; Slice; Source; Structure; Synapses; Synaptic Vesicles; design; millisecond; motor learning; neurophysiology; neurotransmission; neurotransmitter release; novel; novel strategies; postsynaptic; ratiometric; receptor; tool

DESCRIPTION (provided by applicant): The neurotransmitter dopamine is pivotal in decision making, habit and motor learning as well as in drug abuse, schizophrenia, attention deficit hyperactive disorder (ADHD), and Parkinson's disease. This joint effort from three NIH-funded laboratories explores fundamental mechanisms of both pre- and post-synaptic dopaminergic neurotransmission using novel tools. The team includes a chemistry laboratory (Sames) that designs and produces the optical probes, a neurophysiology laboratory that characterizes and analyzes the synaptic release of dopamine using optical recordings at synaptic level (Sulzer), and a molecular neuroscience laboratory that uses a variety of optical approaches to elucidate the structure, activation, and pharmacology of dopamine receptors (Javitch). The three groups have been collaborating intensively for the past five years, and this application results from these collaborations. The novel approaches described here use the first optical methods developed to directly visualize neurotransmitter release, using optical probes known as fluorescent false neurotransmitters (FFNs). The team has now developed a first generation of ratiometric neurotransmitter probes that provide a means to measure pH in synaptic vesicles within neuronal terminals. The probes have also been adapted to simultaneously measure neurotransmission and synaptic vesicle fusion at individual synaptic terminals. At the postsynaptic side, additional probes promise to measure receptor occupancy at unprecedented temporal resolution. While our current data demonstrate the utility of this new generation of optical probes using multiphoton imaging in the acute brain slice, the millisecond-scale events involved in neurotransmission and receptor and transporter activation require virtually simultaneous excitation at two wavelengths, thus necessitating two light sources. We therefore outline how a multiphoton laser will enable multiple and important enhancements to the research of this multidisciplinary group of research teams. Aim 1 outlines how synaptic vesicle fusion and neurotransmitter release will be recorded simultaneously Aim 2 outlines how synaptic vesicle pH will be determined in living synaptic terminals. Aim 3 outlines how caged dopamine used with fluorescence energy transfer will determine the structure, function, and pharmacology of D2 dopamine receptors at unprecedented kinetic resolution.

描述(由申请人提供)：神经递质多巴胺在决策、习惯和运动学习以及药物滥用、精神分裂症、注意力缺陷多动障碍(ADHD)和帕金森病中起关键作用。这项由美国国立卫生研究院资助的三个实验室联合开展的工作，利用新的工具探索突触前和突触后多巴胺能神经传递的基本机制。该团队包括一个设计和生产光学探针的化学实验室(SAMS)，一个使用突触水平上的光学记录来表征和分析多巴胺突触释放的神经生理学实验室(Sulzer)，以及一个使用各种光学方法来阐明多巴胺受体的结构、激活和药理学的分子神经科学实验室(Javitch)。在过去的五年里，这三个小组一直在密集合作，而这一申请就是这些合作的结果。这里描述的新方法使用了第一种光学方法来直接可视化神经递质的释放，使用被称为荧光假神经递质(FFN)的光学探针。该团队现在已经开发出第一代比率神经递质探针，提供了一种测量神经元终末内突触小泡pH的方法。这些探针还被改装成同时测量单个突触终末的神经传递和突触小泡融合。在突触后一侧，更多的探测器有望以前所未有的时间分辨率测量受体的占有率。虽然我们目前的数据证明了这种使用多光子成像的新一代光学探针在急性脑片中的实用性，但涉及神经传递以及受体和转运体激活的毫秒级事件需要在两个波长上几乎同时激发，因此需要两个光源。因此，我们概述了多光子激光将如何使这一多学科研究小组的研究得到多重和重要的增强。目标1概述了如何同时记录突触小泡融合和神经递质释放。目标2概述了如何在活的突触终末中确定突触小泡的pH。目的3概述笼式多巴胺如何与荧光能量转移一起，以前所未有的动力学分辨率决定D2多巴胺受体的结构、功能和药理。