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，注意力缺陷多动症（ADHD）和帕金森氏病中至关重要。这三个由NIH资助的实验室的共同努力探讨了使用新工具使用新工具的前和突触后多巴胺能神经传递的基本机制。该团队包括设计和生产光学探针的化学实验室（SAME），该实验室是一种神经生理学实验室，该实验室表征和分析了使用突触水平（Sulzer）的光学记录（Sulzer）和分子神经科学实验室的突触释放多巴胺的突触释放，并且使用了多种光学方法来使用各种光学方法来阐明结构，而结构，药学，Joav，jovication，dopam incration，dopam acration，dopam。在过去的五年中，这三个小组一直在密集进行协作，这一应用程序来自这些合作。此处描述的新方法使用开发的第一种光学方法直接可视化神经递质释放，使用称为荧光假神经递质（FFN）的光学探针。该团队现在已经开发了第一代比率神经递质探针，该探针提供了一种测量神经元末端突触囊泡的pH值的方法。这些探针也已适应同时测量单个突触终端处的神经传递和突触囊泡融合。在突触后一侧，其他探针有望在空前的时间分辨率下测量受体占用。虽然我们当前的数据证明了这种新一代光学探针在急性脑切片中使用多光子成像的实用性，但涉及神经传递的毫秒尺度事件，受体和转运蛋白激活几乎需要在两个波长上同时激发，因此需要两个光源。因此，我们概述了多光子激光器将如何为该多学科研究团队的研究启用多个重要的增强功能。 AIM 1概述了如何同时记录突触囊泡融合和神经递质释放的AIM 2概述如何在活体突触末端确定突触囊泡pH。 AIM 3概述了如何在空前的动力学分辨率下确定D2多巴胺受体的结构，功能和药理学的结构，功能和药理学。