Optical tools for extended neural silencing

用于延长神经沉默的光学工具

• 批准号: 9055530
• 负责人: Matthew J Kennedy
• 金额: $ 23.19万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2017-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9055530
• 关键词: Acute; Adopted; Adoption; Anatomy; Animals; Behavior; Behavioral; Biosensor; Bontoxilysin; Brain; Brain region; Cells; Chloride Ion; Chlorides; Clostridium; Communities; Complement; Complex; Coupled; DNA; Designer Drugs; Dimerization; Directed Molecular Evolution; Disease; Engineering; G-Protein Signaling Pathway; GABA Receptor; Genetic Recombination; Genetic Transcription; Goals; Halorhodopsins; Hippocampus (Brain); Hour; Individual; Label; Lesion; Light; Maps; Mediating; Neurons; Neurosciences; Neurotoxins; Neurotransmitters; Operative Surgical Procedures; Opsin; Optical reporter; Optics; Peptide Hydrolases; Phototoxicity; Physiologic pulse; Population; Presynaptic Terminals; Process; Property; Protein Fragment; Proteins; Proton Pump; Reagent; Receptor Activation; Regulation; Research Personnel; Resolution; Rodent; Serotyping; Signal Transduction; Slice; Specificity; Synapses; System; Technology; Testing; Time; Toxicity Tests; Toxin; Training; Update; Viral; Work; Yeasts; base; cell type; cohort; in vivo; insight; microbial; millisecond; neural circuit; neuroregulation; neurotransmission; neurotransmitter release; novel strategies; optogenetics; public health relevance; receptor; reconstitution; relating to nervous system; spatiotemporal; temporal measurement; tetanospasmin; tool; transmission process; vector

 DESCRIPTION: Conditionally silencing the activity of specific neural ensembles is a powerful approach for mapping the circuits responsible for specific behaviors. While microbial opsin tools currently exist to silence neural activity with light, these tools have significant limitations tha make them unsuitable for applications that require persistent silencing over the course of minutes or hours. Longer term neuronal silencing has been classically achieved by expressing the catalytic light chain of Clostridium neurotoxins, which disrupt neurotransmitter release, however this approach suffers from poor temporal and spatial control. Recent chemogenetic approaches, including designer receptors exclusively activated by designer drugs (DREADDs) have been developed for persistent silencing, but lack the spatiotemporal benefits of optogenetic approaches. Thus, there remains an unmet need for silencing tools that combine the robust and persistent silencing qualities of Clostridium neurotoxin and chemogenetic approaches, with the spatiotemporal control of optogenetics. Here we will develop a completely new toolkit for rapid, extended neural silencing with a single, brief light pulse that can be readiy activated by either single or multiphoton excitation for in vivo systems. Our tools will not only b applicable to neurotransmitter systems, but also to neuromodulatory systems and glial transmission. These powerful tools will complement and extend the existing opto- and chemogenetic silencing toolset by allowing rapid, robust and persistent silencing with a single light pulse. Because our strategy operates on a completely different principle than current tools, it could be multiplexed with existing approaches for complex remote control of circuit activity during behavior.

 产品说明：抑制特定神经系统的活动是一种强大的方法来映射负责特定行为的电路。虽然目前存在微生物视蛋白工具来用光使神经活动沉默，但这些工具具有显著的局限性，使得它们不适合需要在数分钟或数小时的过程中持续沉默的应用。较长期的神经元沉默已经经典地通过表达梭菌神经毒素的催化轻链来实现，其破坏神经递质释放，然而这种方法受到时间和空间控制差的影响。最近的化学遗传学方法，包括专门由设计药物激活的设计受体（DREADD）已经被开发用于持续沉默，但缺乏光遗传学方法的时空益处。因此，对于将梭菌神经毒素和化学遗传方法的稳健和持久沉默品质与光遗传学的时空控制相结合的沉默工具仍然存在未满足的需求。在这里，我们将开发一种全新的工具包，用于快速，延长神经沉默，使用单个短暂的光脉冲，可以通过单光子或多光子激发激活体内系统。我们的工具不仅适用于神经递质系统，而且适用于神经调节系统和胶质传递。这些强大的工具将补充和扩展现有的光学和化学基因沉默工具集，通过使用单个光脉冲实现快速，稳健和持久的沉默。由于我们的策略与当前工具的原理完全不同，因此它可以与现有方法复用，用于在行为期间对电路活动进行复杂的远程控制。