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

• 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
• 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

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 -在解剖神经电路、细胞信号和操纵方面有着巨大的前景 清醒的、行为正常的动物的神经生理系统。然而，实施的技术限制 在清醒、自由运动行为中解剖神经调节器的光遗传学在与 需要对受体信号进行离散时空控制的范例，以及在研究神经时 具有非常小的多样的、“难以到达”的结构的电路，例如不同种类的大脑核团。至 使用神经药物受体底物，几乎每个领域的神经科学家都使用插管(简单 金属管)，以及最近采用系留光纤用于体内光遗传学以控制局部 神经调节剂单胺或神经肽的释放。不幸的是，这些目前的方法相当于 有限且难以实现，因为它们严重限制了对受体信号的时空控制 离散细胞类型中的通路。此外，目前的技术缺乏一个完整的工具箱，用于多路、亚蜂窝、 神经调节剂的主要手段--G蛋白偶联受体信号的时空调控 大脑中的交流。出于这些原因，一项将神经科学与生物化学相结合的创新努力 而药理学是必要的，以便在体外和体内对gpr进行时空控制。 神经调节剂信号。因此，我们在这里直接讨论RFA-NS-16-775的核心目标 循规蹈矩。这项提议的中心目标是开发一种尖端的v2.0 Opto-XR受体， 在体外和自由活动的动物中，在空间和时间上控制神经调节剂信号。我们有 提出了一种独特的综合方法来实现这一目标，它将药理学家、生理学家、 生物化学家和神经学家以一种独特的平行方式聚集在一起。在我们将制定的两个具体目标中 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-功能分析和 深入的体外鉴定；和2)开发实用性和鉴定Gi和Gs版本的Opto-XR V2.0使用传统和无线光生技术在活体和自由移动行为模型中构建 接近了。该提案的成功完成将为更广泛的神经科学界提供 期待已久的GPCRs时空操作-清醒状态下神经回路内的神经调制信号 行为自由的动物。这项新技术还将进一步拓宽有能力的方法的领域 对脑组织中神经调节器功能的离散控制和光动力学模拟。