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

项目摘要 在这里，我们建议开发一种新的狂犬病病毒为基础的战略，表达两个通道视紫红质蛋白 独立操纵兴奋性和抑制性跨膜电位在同一人口的 神经元结合辅助病毒和狂犬病假型中细胞类型特异性启动子的使用， 一套新的病毒将能够完全控制特定神经网络亚型的兴奋和抑制， 非转基因动物我们将在这里应用新技术，以确定细胞类型的具体贡献， 前馈皮层整合对复杂视觉特征选择性的出现， 动物模型而我们的目标主要是发展这种新的方法来研究结构-功能组织 这种技术将很容易适用于研究任何大脑区域。 我们提出的技术结合了最近可用的通道视紫红质阳离子和阴离子通道， 我们最近开发的病毒策略，用于访问非- 转基因物种（Liu等，2013年，Curr Biol）为前所未有的精细规模研究打开了大门， 高度视觉哺乳动物的结构-功能关系。对于本提案，我们将采用这些新策略 为了能够光遗传学操纵涉及高阶视皮层感受野的特定细胞类型， 阵这些研究将代表对抑制性神经元和潜在神经元的最直接的体内评估。 大型高度视觉哺乳动物的层内和层间电路，促进我们对基本原理的理解 视觉过程产生并依赖于复杂的皮质结构。

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.