In vivo Technology for Fast Optical Control of Neural Circuits

了解脑细胞如何产生复杂的过程，如行动、思想和情感，受到目前可用的控制神经组织中特定细胞的工具的限制。对脑细胞进行快速的基因定向控制将有助于阐明特定神经元类型的作用，并极大地促进我们对大脑功能的理解。为了能够精确地扰动活的电路，光激活离子通道，称为通道视紫红质-2 (ChR2)，将用于基因靶向，毫秒级的神经元光学激发。另一种被称为NpHR的蛋白质已经被确定为对神经活动的时间精确的光学抑制。这两种蛋白质在哺乳动物神经元中起作用，共同形成一个完整的，互补的系统，用于多模式，高速，遗传靶向，全光学的活神经回路研究。该项目包括广泛的技术开发、测试和传播计划，将为生理学、神经科学和生物医学工程界提供通用的、强大的工具。具体而言，将开发用于活体动物的高速光学成像试剂和同时成像和光刺激/抑制的技术。这是一种全新的方法，可能会彻底改变在完整神经系统中探测回路的方式。这些工具具有广泛的应用范围，这项工作的结果可能是通用的，足以让研究人员确定细胞类型选择的贡献，并将其与几乎任何感兴趣的大脑区域的电路动力学和行为联系起来。这些新技术也为学生提供了独特的培训机会，他们将转到其他机构并传递专业知识，多年来，一代又一代的科学家和工程师骨干接受了这项强大技术的强化培训。

Understanding how brain cells give rise to complex processes like action, thought, and emotion is limited by the current availability of tools to control specific cells within neural tissue. Fast, genetically targeted control over brain cells would allow elucidation of the role of specific neuronal types, and greatly advance our understanding of brain function. To enable precise perturbation of living circuits, the light-activated ion channel, called channelrhodopsin-2 (ChR2), will be used for genetically targeted, millisecond-timescale optical excitation of neurons. Another protein, called NpHR, has been identified for temporally-precise optical inhibition of neural activity. Both proteins function in mammalian neurons, together forming a complete, complementary system for multimodal, high-speed, genetically-targeted, all-optical investigation of living neural circuits. This project encompasses a broad technology development, testing, and dissemination plan that will yield generalizable, powerful tools for the physiology, neuroscience and biomedical engineering communities. Specifically, high-speed optical imaging reagents and technology for simultaneous imaging and optical stimulation/inhibition will be developed, in living animals. This is a fundamentally novel approach that could revolutionize the way circuits are probed in intact neural systems. These tools have an enormous range of applications, and the outcome of this work may be versatile enough to allow investigators to determine the contribution of a cell type of choice and relate it to circuit dynamics and behavior in virtually any brain region of interest. These new technologies also afford unique training opportunities for students, who will move on to other institutions and pass on expertise, leading over years to generation of a widespread cadre of scientists and engineers with intensive training in this powerful technology.