McGovern Institute for Brain Research and Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA.
Nat Protoc. 2010 Feb;5(2):247-54. doi: 10.1038/nprot.2009.228. Epub 2010 Jan 21.
A major long-term goal of systems neuroscience is to identify the different roles of neural subtypes in brain circuit function. The ability to causally manipulate selective cell types is critical to meeting this goal. This protocol describes techniques for optically stimulating specific populations of excitatory neurons and inhibitory interneurons in vivo in combination with electrophysiology. Cell type selectivity is obtained using Cre-dependent expression of the light-activated channel Channelrhodopsin-2. We also describe approaches for minimizing optical interference with simultaneous extracellular and intracellular recording. These optogenetic techniques provide a spatially and temporally precise means of studying neural activity in the intact brain and allow a detailed examination of the effect of evoked activity on the surrounding local neural network. Injection of viral vectors requires 30-45 min, and in vivo electrophysiology with optogenetic stimulation requires 1-4 h.
系统神经科学的一个主要长期目标是确定神经亚型在大脑回路功能中的不同作用。能够有因果地操纵选择性细胞类型对于实现这一目标至关重要。本协议描述了在体内结合电生理学对特定群体的兴奋性神经元和抑制性中间神经元进行光刺激的技术。细胞类型选择性是通过 Cre 依赖性表达光激活通道 Channelrhodopsin-2 获得的。我们还描述了最大限度地减少光学干扰与同时进行的细胞外和细胞内记录的方法。这些光遗传学技术为研究完整大脑中的神经活动提供了一种空间和时间上精确的方法，并允许对诱发活动对周围局部神经网络的影响进行详细检查。病毒载体的注射需要 30-45 分钟，而带有光遗传学刺激的体内电生理学需要 1-4 小时。
