An optogenetics- and imaging-assisted simultaneous multiple patch-clamp recording system for decoding complex neural circuits.
An optogenetics- and imaging-assisted simultaneous multiple patch-clamp recording system for decoding complex neural circuits.
复制标题
DOI：
10.1038/nprot.2015.019
复制
发表时间：
2015-03
期刊：
影响因子：
14.8
通讯作者：
Zhu JJ
中科院分区：
文献类型：
--
作者：
Wang G;Wyskiel DR;Yang W;Wang Y;Milbern LC;Lalanne T;Jiang X;Shen Y;Sun QQ;Zhu JJ
破译神经回路是理解大脑功能和功能障碍的核心，但这仍然是一项艰巨的任务。为了便于解剖神经元回路，这一过程需要突触连接的功能分析和相互连接的神经元的形态识别，我们在这里提出了一种稳定的八元片钳同步记录方法。该方法允许对同时记录的4-8个神经元和/或顺序记录的10-30个神经元之间的突触相互连接进行生理分析，并允许对记录的中间神经元的>进行解剖鉴定85%，对记录的主神经元的>进行解剖鉴定99%。我们描述了如何将该方法应用于啮齿动物组织切片；然而，它可以用于其他模式生物。我们还描述了机械，电子，光学和软件程序的最新改进和优化，这些程序对于实现基于单光子和双光子显微镜的组合，光遗传学和成像辅助，稳定，同时四倍视点膜片钳记录系统至关重要。从开始的仪器组装和软件安装到全面运行，系统的设置可在3-4天内完成。
Deciphering neuronal circuitry is central to understanding brain function and dysfunction, yet it remains a daunting task. To facilitate the dissection of neuronal circuits, a process requiring functional analysis of synaptic connections and morphological identification of interconnected neurons, we present here a method for stable simultaneous octuple patch-clamp recordings. This method allows physiological analysis of synaptic interconnections among 4–8 simultaneously recorded neurons and/or 10–30 sequentially recorded neurons, and it allows anatomical identification of >85% of recorded interneurons and >99% of recorded principal neurons. We describe how to apply the method to rodent tissue slices; however, it can be used on other model organisms. We also describe the latest refinements and optimizations of mechanics, electronics, optics and software programs that are central to the realization of a combined single- and two-photon microscopy–based, optogenetics- and imaging-assisted, stable, simultaneous quadruple–viguple patch-clamp recording system. Setting up the system, from the beginning of instrument assembly and software installation to full operation, can be completed in 3–4 d.