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

下一代 Opto-GPCR：神经调节信号传导的时空模拟

• 批准号: 9815886
• 负责人: Michael R. Bruchas
• 金额: $ 103.87万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-11-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9815886
• 关键词: Next; gen; Opto; GPCRs; spatiotemporal

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 - 使用光吸引生物系统 - 对神经回路，细胞信号传导和操纵的解剖有很大的希望 神经生理系统在清醒中，行为动物。但是，实施的技术限制 在使用神经调节剂时，在使用神经调节剂时，自由移动行为很明显 需要离散的接收器信号传导的空间控制以及研究中性时的范例 潜水员非常小的电路“难以触及”的结构，例如异质脑核。到 在几乎每个田间都使用神经药物受体，神经科学家使用套管（简单 金属管），最近采用了用于体内光遗传学的束带光纤，以控制局部 神经调节剂单胺或神经肽的释放。不幸的是，这些当前的方法是 有限且难以实施，因为它们严重限制了对受体信号的时空控制 离散单元类型中的途径。此外，当前的技术缺乏用于多路复用，亚细胞的完整工具盒 G蛋白耦合受体信号传导的时空控制，神经调节剂的主要平均值 大脑的交流。由于这些原因，将神经科学与生物化学相结合的创新努力 为了在体外和体内控制GPCR-的时空，需要药理学。 神经调节器信号传导。因此，在这里，我们直接解决了此RFA-NS-16-775的核心目标 以下方式。该提案的主要目标是开发一个尖端的v2.0 opto-xr接收器 在空间和临时控制体外和自由移动动物的神经调节剂信号传导。我们有 提出了一种独特的综合方法，以实现这一目标，该目标带来了药物，生理学家， 生物化学家和神经科学家以独特的平行方式一起。在两个具体目标中，我们将发展 并在体外和体内测试这些新型工具：1）开发具有带有opo-xr v2.0受体的突变体GI和GS 使用结构功能分析和 老实说，体外表征； 2）开发效用并表征opo-xr的GI和GS版本 v2.0在体内以及使用传统和无线光学遗传学的自由移动行为模型中构造 方法。该提案的成功完成将为更广泛的神经科学社区提供 长期等待的GPCR的时空操纵 - 神经通路内神经调节剂信号传导 自由表现动物。这项新技术还将进一步宽图范围 神经调节剂功能在脑组织中的离散控制和光学模拟。

项目成果

Optogenetic approaches for dissecting neuromodulation and GPCR signaling in neural circuits.

• DOI: 10.1016/j.coph.2016.11.001
• 发表时间: 2017-02
• 期刊: Current opinion in pharmacology
• 影响因子: 4
• 作者: Spangler SM;Bruchas MR
• 通讯作者: Bruchas MR