Anion channelrhodopsin-based viral tools to manipulate brain networks in behaving animals

基于阴离子通道视紫红质的病毒工具可操纵行为动物的大脑网络

基本信息

批准号：
9321918
负责人：
VALENTIN DRAGOI
金额：
$ 95.18万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-21 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9321918
关键词：
Address Algae Animals Anions Behavior Brain Cations Cells Cloning Cognitive Color Complex Decision Making Disease Dissection Effectiveness Electrophysiology (science)Exhibits Gene Delivery Generations Genes Glutamates Goals Human In Situ Individual Injectable Injection of therapeutic agent Kinetics Light Lighting Macaca mulatta Mammals Membrane Potentials Mental disorders Methods Modeling Molecular Molecular Profiling Monkeys Mus Neurons Neurosciences Opsin Optics Pattern Photosensitivity Physiological Population Primates Procedures Process Production Property Protein Engineering Protocols documentation Psyche structure Recovery Research Research Project Grants Rhodopsin Role Scientist Sensory Sodium Specificity System Techniques Testing Viral Viral Vector Visual area striata base behavior influence brain tissue cell type cognitive function cognitive process excitatory neuron expression vector extracellular improved information processing inhibitory neuron interest mouse model nervous system disorder neural circuit neuronal survival nonhuman primate novel optogenetics patch clamp promoter public health relevance rate of change relating to nervous system response selective expression tool vector

项目摘要

DESCRIPTION (provided by applicant): Examining neural circuits crucially relies on the ability to activate or silence individual circuit components to subsequently assess their impact on other parts of the circuit and their influence on behavior. Recent refinements of viral tools for gene delivery have allowed optogenetic methods to target cells based on specific cell types, localization, and connectivity. The physiological dissection of targeted circuits has been extremely successful in the mouse brain, but remains of limited use in non-human primate brain. We plan to develop and test a new generation of viral tools that will allow us to both activate and suppress different cell types in non-human primate models. To accomplish our aims we have assembled an expert team with complementary expertise composed of a biochemist and photobiologist (John Spudich), a molecular neuroscientist (Roger Janz), and a systems and computational neuroscientist (Valentin Dragoi). Our approach builds upon recently discovered anion-conducting channelrhodopsins (ACRs), which perform with perfect anion selectivity, photosensitivity orders of magnitude greater than current optogenetic rhodopsins, and enable highly efficient neuron hyperpolarization. We believe that our ACR constructs will open a new chapter in targeted neuro-suppression. In addition, we will use new neuron-activating (depolarizing) cation-conducting channelrhodopsins (CCRs) that have ~3-fold greater unitary conductance, faster recovery from excitation, and higher sodium selectivity than the commonly used channelrhodopsin-2. We will construct viral vectors encoding ACR-CCR pairs and, using spectrally different ACRs, ACR-ACR pairs, enabling efficient wavelength-selected neuron activation or suppression in large populations. The effectiveness of these viral vectors will be tested in cultured and in situ mouse neurons and in the primary visual cortex (V1) of behaving monkeys. Developing these powerful tools will be invaluable for probing neural circuits in non-human primate models, finally allowing the interrogation of microcircuits underlying primate cognitive function.

描述（由申请人提供）：检查神经回路主要依赖于激活或沉默单个回路组件的能力，以随后评估它们对回路其他部分的影响以及它们对行为的影响，最近对基因传递病毒工具的改进已经允许光遗传学。基于特定细胞类型、定位和连接性的目标细胞递送方法在小鼠大脑中非常成功，但在非人类灵长类动物大脑中的应用仍然有限。新一代病毒工具将使我们能够激活和为了实现我们的目标，我们组建了一个具有互补专业知识的专家团队，由生物化学家和光生物学家 (John Spudich)、分子神经科学家 (Roger Janz) 以及系统和计算神经科学家 (Roger Janz) 组成。 Valentin Dragoi）。我们的方法建立在最近发现的阴离子传导通道视紫红质（ACR）的基础上，它具有完美的阴离子选择性，光敏度比现有的高几个数量级。我们相信，我们的 ACR 构建体将开启靶向神经抑制的新篇章。此外，我们将使用具有新的神经元激活（去极化）阳离子传导通道视紫红质（CCR）。比常用的视紫红质通道蛋白-2 具有大约 3 倍的单位电导、更快的激发恢复和更高的钠选择性。 ACR-CCR 对，以及使用光谱不同的 ACR、ACR-ACR 对，可以在大量群体中实现有效的波长选择神经元激活或抑制。这些病毒载体的有效性将在培养和原位小鼠神经元以及初级视觉中进行测试。开发这些强大的工具对于探测非人类灵长类动物模型中的神经回路非常有价值，最终可以探究灵长类动物认知功能的微回路。