DEVELOPMENT OF A FOS-CHANNEL RHODOPSIN TRANSGENIC MOUSE

描述(由申请人提供)：即刻早期基因c-fos将神经活动与基因表达结合在一起，已被广泛用作识别刺激特异性神经集合的标记。在这里，我们建议创建和描述使用c-fos启动子来驱动光激活的阳离子通道通道视紫红质(ChR)与黄色荧光蛋白YFP偶联的转基因小鼠。这项提议融合了尖端技术(直接可视化基因表达和使用光激活离子通道重新激活电路)，促进了对系统神经科学中新问题的研究。FOS-CHR转基因小鼠将允许我们1)标记激活细胞的身份，2)随后驱动这些神经元的活动，以测试特定神经元在感知和行为中的作用。神经元亚群的受控重新激活将回答有关特定神经元亚群如何编码感知和行为的重要问题，并可能促进对已被药物暴露或疾病改变的适应不良神经回路的积极再训练。与公共健康相关：哺乳动物的大脑包含数百万到数万亿个神经元，这些神经元驱动着一系列复杂的行为。要了解特定的神经元群是如何参与并编码感知和行为的，需要对这些神经子集进行识别和分析。即刻早期基因c-fos的表达可以“标记”任务或刺激特定神经元的群体，为定义神经亚群提供了一个功能标准。利用FOS基因启动子序列驱动光激活的阳离子通道通道视紫红质(ChR)的表达，我们将创建并鉴定FOS-CHR转基因小鼠。这些动物将使我们能够识别并重新激活功能定义的神经集合，以了解受控的小神经元子集的重新激活如何驱动感知和行为。此外，神经重新激活可能允许重新训练特定的回路，以消除成瘾或精神障碍中的不良适应行为。

DESCRIPTION (provided by applicant): The immediate early gene c-fos couples neural activity to gene expression and has been widely used as a marker to identify stimulus-specific neural ensembles. Here we propose to create and characterize transgenic mice that use the c-fos promoter to drive expression of the light-activated cation channel, channelrhodopsin (ChR), coupled to the yellow-fluorescent protein YFP. This proposal presents a fusion of cutting-edge technologies (direct visualization of gene expression and use of light-activated ion channels for circuit reactivation) that facilitate investigation of novel questions in systems neuroscience. Fos-ChR transgenic mice will allow us to 1) mark the identity of activated cells and 2) subsequently drive activity in these neurons to test the role of specific neurons in perception and behavior. Controlled reactivation of neuronal subsets will answer important questions about how specific neuronal subsets encode perception and behavior, and may facilitate active retraining of maladaptive neural circuits that have been altered by drug exposure or disease. PUBLIC HEALTH RELEVANCE: The mammalian brain contains millions to trillions of neurons that drive a wide array of complex behaviors. Understanding how specific neuronal ensembles are engaged by and encode perception and behavior requires identification and analysis of these neural subsets. Expression of the immediate-early gene c-fos can "mark" populations of task- or stimulus-specific neurons, providing a functional criterion to define neural subpopulations. Using fos gene promoter sequences to drive expression of the light-activated cation channel, channelrhodopsin, (ChR) we will create and characterize fos-ChR transgenic mice. These animals will enable us to identify and then reactivate functionally-defined neural ensembles to understand how the controlled reactivation of small subsets of neurons can drive perception and behavior. In addition, neural reactivation may allow retraining of specific circuits to eliminate maladaptive behaviors in addiction or psychiatric disorders.