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（由适用提供）：检查中性电路完全取决于激活或沉默单个电路组件的能力，以评估其对电路其他部位的影响及其对行为的影响。用于基因输送的病毒工具的最新改进允许基于特定细胞类型，定位和连通性靶向细胞的光遗传学方法。靶向电路的物理解剖在小鼠大脑中非常成功，但在非人类灵长类动物大脑中的使用有限。我们计划开发和测试新一代的病毒工具，使我们同时激活和在非人类私有模型中抑制不同的细胞类型。为了实现我们的目标，我们将一支专家团队组成了一位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（Valentin Dragoi）的完整专家组成。我们的方法建立在最近发现的阴离子传导通道旋转蛋白（ACR）的基础上，它们以完美的阴离子选择性，光敏度的数量级高于当前的光遗传学旋转蛋白，并启用高效的神经元过度溶质化。我们认为，我们的ACR结构将在有针对性的神经抑制中开设新篇章。此外，我们还将使用新的神经激活（去偏基化）阳离子传导通道旋转蛋白（CCRS），其单位电导率大约比常见的通道Rhopopsin-2高约3倍，更快地从兴奋中恢复和更高的钠选择性。我们将构建编码ACR-CCR对的病毒矢量，并使用频谱ACR-ACR-ACR对，从而在大型种群中启用有效的波长选择的神经元激活或抑制。这些病毒载体的有效性将在培养和原位小鼠神经元以及行为猴子的主要视觉皮层（V1）中进行测试。开发这些强大的工具对于在非人类灵长类动物模型中探测神经元将是无价的，最终允许对灵长类动物认知功能的微电路进行审问。