Viral strategies for bi-directional optogenetic control of specific cell types in neocortex of non-transgenic animals

非转基因动物新皮质中特定细胞类型双向光遗传学控制的病毒策略

• 批准号: 10057749
• 负责人: David C Lyon
• 金额: $ 43.18万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057749
• 关键词: Afferent Neurons; Animal Model; Anions; Brain region; Cations; Cells; Characteristics; Complex; Effectiveness; Electrophysiology (science); Felis catus; Glycoproteins; Goals; Helper Viruses; High Frequency Oscillation; Image; Individual; Infection; Injections; Ion Channel Gating; Lasers; Mammals; Membrane Potentials; Methods; Modeling; Neocortex; Nervous system structure; Neural Network (Anatomic); Neurons; Opsin; Output; Pattern; Physiologic pulse; Play; Population; Property; Proteins; Rabies; Rabies virus; Rattus; Reporter; Research; Role; Signal Transduction; Stimulus; Structure; Structure-Activity Relationship; System; Systems Analysis; Techniques; Testing; Time; Training; Tva receptor; V2 neuron; Validation; Variant; Viral; Virus; Visual; Visual Cortex; area striata; base; cell type; design; efficacy testing; fluorescence imaging; in vivo; inhibitory neuron; light gated; millisecond; neural network; neurophysiology; novel; novel strategies; optical imaging; optogenetics; presynaptic; promoter; receptive field; red fluorescent protein; visual process

Project Summary Here we propose to develop a novel rabies virus based strategy to express two channelrhodopsin proteins independent manipulation of excitatory and inhibitory transmembrane potentials in the same population of neurons. Combined with the use of cell-type specific promoters in helper viruses and rabies pseudotyping, the new suite of viruses will enable full control of excitation and inhibition over specific neural network subtypes of non-transgenic animals. We will apply the new technique here to determine cell type specific contributions of feedforward cortical integration on the emergence of complex visual feature selectivity using highly visual animal models. While our goal is primarily to develop this novel method to study structure-function organization of visual cortex, this technique will be readily applicable to study any brain region. Our proposed technique combines recently available channelrhodopsin cation and anion channel with our recently our developed viral strategies for accessing specific cell types and specific circuits in non- transgenic species (Liu et al., 2013, Curr Biol) opening the door for unprecedented fine scale study of structure-function relationships in highly visual mammals. For this proposal we will apply these new strategies to enable optogenetic manipulation of specific cell types involved in higher order visual cortex receptive field formation. These studies will represent the most direct in vivo assessment of inhibitory neurons and underlying intra- and inter-laminar circuitry of a large, highly visual mammal, advancing our understanding of how basic visual processes arise and depend on complex cortical structure.

项目摘要 在这里，我们建议开发一种基于狂犬病病毒的新型策略来表达两种通道旋转蛋白 在同一人群中独立操纵兴奋性和抑制性跨膜电位 神经元。结合使用在辅助病毒和狂犬病假型中使用细胞类型的特定启动子， 新的病毒套件将使人们能够完全控制对特定神经网络亚型的激发和抑制 非转基因动物。我们将在此处应用新技术来确定细胞类型的特定贡献 使用高度视觉 动物模型。虽然我们的目标主要是开发这种新颖的方法来研究结构功能组织 在视觉皮层中，该技术将很容易适用于研究任何大脑区域。 我们提出的技术将最近可用的ChannelRhopopsin阳离子和阴离子通道与 我们最近开发的病毒策略，用于访问非 - 非特定细胞类型和特定电路 转基因物种（Liu等人，2013年，Curr Biol）为前所未有的细节研究开门 高度视觉哺乳动物中的结构功能关系。对于此建议，我们将应用这些新策略 为了使涉及高阶视觉皮层接受场的特定细胞类型的光遗传操作 形成。这些研究将代表抑制性神经元和基础的最直接的体内评估 大型，高视觉哺乳动物的层间和层间电路，促进了我们对基础的理解 视觉过程发生并取决于复杂的皮质结构。