Next-gen Opto-GPCRs: spatiotemporal simulation of neuromodulator signaling

下一代 Opto-GPCR：神经调节信号传导的时空模拟

基本信息

批准号：
9213972
负责人：
Michael R. Bruchas
金额：
$ 110.38万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-09 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9213972
关键词：
Action Potentials Activities of Daily Living Address Adopted Adoption Animal Model Animals Appetitive Behavior Architecture Arrestins Behavior Behavioral Biochemical Biochemistry Biology Boxing Brain Brain Diseases Cannulas Cell Nucleus Cells Clinical Color Communication Communities Complex Corticotropin-Releasing Hormone Dissection Dopamine Engineering Fiber Optics G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GTP-Binding Proteins Generations Genetic Goals Health Human Image In Vitro Ion Channel Ion Pumps Ions Knowledge Ligands Light Maps Mediating Mental disorders Metals Methods Modeling Molecular Monitor Morphologic artifacts Mus Neurobiology Neurodegenerative Disorders Neuromodulator Neurons Neuropeptides Neurosciences Norepinephrine Opioid Opsin Organism Pharmacology Physiological Processes Physiology Process Pump Receptor Signaling Resolution Series Serotonin Signal Pathway Signal Transduction Slice Structure System Techniques Technology Testing Time Translating Tube Variant Viral Wireless Technology Work abstracting awake base biological systems brain tissue cell type design designer receptors exclusively activated by designer drugs hypocretin in vivo innovation monoamine multidisciplinary mutant neural circuit neurophysiology neuroregulation neurotechnology new technology next generation novel optogenetics peptide G receptor relating to nervous system research study simulation spatiotemporal tool

项目摘要

Project Summary/Abstract: The emerging field of optogenetics — using light to engage biological systems — holds tremendous promise for dissection of neural circuits, cellular signaling and manipulating neurophysiological systems in awake, behaving animals. However, the technological limits for implementing optogenetics in dissecting neuromodulators in awake, freely-moving behavior is clear while working with paradigms that require discrete spatiotemporal control of receptor signaling and when investigating neural circuits that have very small diverse, “hard to reach” architecture, such as heterogeneous brain nuclei. To engage neuropharmacological receptor substrates, neuroscientists in nearly every field use cannulas (simple metal tubes) and have more recently adopted tethered fiber optics for in vivo optogenetics to control local release of neuromodulator monoamine or neuropeptides. Unfortunately, these current methods are rather limited and difficult to implement because they severely limit the spatiotemporal control over receptor signaling pathways in discrete cell types. Moreover, current technology lacks a full tool box for multiplexed, subcellular, spatiotemporal control of G protein coupled receptor signaling, the predominant means for neuromodulator communication in the brain. For these reasons, an innovative effort combining neuroscience with biochemistry and pharmacology was necessary in order to bring spatial-temporal in vitro and in vivo control over GPCR- neuromodulator signaling. Therefore, here we directly address the central goals of this RFA-NS-16-775 in the following manner. The central goal of this proposal is to develop a cutting-edge v2.0 Opto-XR receptors that spatially and temporally control neuromodulator signaling in vitro and in freely moving animals. We have proposed an uniquely integrated approach to achieve this goal that brings pharmacologists, physiologists, biochemists, and neuroscientists together in a unique parallel manner. In the two specific aims we will develop and test these novel tools in vitro and in vivo: 1) To develop mutant Gi and Gs, Opto-XR v2.0 receptors with greater signaling dynamics and altered color spectra and sensitivity using structure-function analyses and thorough in vitro characterization; and 2) To develop utility and characterize Gi and Gs versions of Opto-XR v2.0 constructs in vivo and in models of freely-moving behavior using both traditional and wireless optogenetic approaches. Successful completion of the proposal will provide the wider community of neuroscience with a long awaited spatiotemporal manipulation of GPCRs – neuromodulator signaling within neural circuits in awake freely behaving animals. This new technology will also further widen the field for approaches that are capable of discrete control and optodynamic simulation of neuromodulator function in brain tissue.

项目摘要/摘要：光遗传学的新兴领域——利用光参与生物系统 - 为神经回路的解剖、细胞信号传导和操纵带来了巨大的希望清醒、行为动物的神经生理系统然而，实施的技术限制。光遗传学在清醒、自由移动行为中解剖神经调节剂的作用是清楚的，同时与需要对受体信号进行离散时空控制以及研究神经系统时的范例具有非常小、多样化、“难以到达”架构的电路，例如异构脑核。涉及神经药理学受体底物，几乎每个领域的神经科学家都使用插管（简单金属管），最近采用系留光纤进行体内光遗传学来控制局部不幸的是，这些当前的方法相当缺乏神经调节剂单胺或神经肽的释放。有限且难以实施，因为它们严重限制了对受体信号传导的时空控制此外，当前的技术缺乏用于多重、亚细胞、 G 蛋白偶联受体信号传导的时空控制，神经调节剂的主要手段由于这些原因，神经科学与生物化学相结合的创新努力。为了在体外和体内对 GPCR 进行时空控制，药理学是必要的因此，在这里我们直接解决 RFA-NS-16-775 中的中心目标。该提案的中心目标是开发一种尖端的 v2.0 Opto-XR 受体。我们已经在体外和自由活动的动物中空间和时间控制神经调节信号。提出了一种独特的综合方法来实现这一目标，该方法使药理学家、生理学家、生物化学家和神经科学家以独特的并行方式共同致力于这两个具体目标。并在体外和体内测试这些新工具：1) 开发突变型 Gi 和 Gs、Opto-XR v2.0 受体使用结构功能分析增强信号动态并改变色谱和灵敏度彻底的体外表征；以及 2) 开发实用性并表征 Opto-XR 的 Gi 和 Gs 版本 v2.0 使用传统和无线光遗传学构建体内和自由移动行为模型该提案的成功完成将为更广泛的神经科学界提供一个机会。期待已久的 GPCR 时空操纵——清醒状态下神经回路内的神经调节信号传导这项新技术也将进一步拓宽有能力的方法的领域。脑组织神经调节功能的离散控制和光动力学模拟。