title: An optogenetics- and imaging-assisted simultaneous multiple patch-clamp recording system for decoding complex neural circuits.
abstract: 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.
authors: Department of Pharmacology, University of Virginia, Charlottesville, Virginia, USA.
authors: Guangfu
authors: G
authors: Wang
authors: Department of Pharmacology, University of Virginia, Charlottesville, Virginia, USA.
authors: Daniel R
authors: DR
authors: Wyskiel
authors: Department of Zoology and Physiology, University of Wyoming, Laramie, Wyoming, USA.
authors: Weiguo
authors: W
authors: Yang
authors: 1] Department of Pharmacology, University of Virginia, Charlottesville, Virginia, USA. [2] Department of Chemistry, University of Virginia, Charlottesville, Virginia, USA.
authors: Yiqing
authors: Y
authors: Wang
authors: Department of Pharmacology, University of Virginia, Charlottesville, Virginia, USA.
authors: Lana C
authors: LC
authors: Milbern
authors: 1] Department of Pharmacology, University of Virginia, Charlottesville, Virginia, USA. [2] Neuroscience Graduate Program, Department of Health and Life Sciences, Bordeaux University, Bordeaux, France.
authors: Txomin
authors: T
authors: Lalanne
authors: Department of Pharmacology, University of Virginia, Charlottesville, Virginia, USA.
authors: Xiaolong
authors: X
authors: Jiang
authors: Department of Neurobiology and Key Laboratory of Medical Neurobiology of Chinese Ministry of Health, Zhejiang University School of Medicine, Hangzhou, China.
authors: Ying
authors: Y
authors: Shen
authors: Department of Zoology and Physiology, University of Wyoming, Laramie, Wyoming, USA.
authors: Qian-Quan
authors: QQ
authors: Sun
authors: 1] Department of Pharmacology, University of Virginia, Charlottesville, Virginia, USA. [2] Department of Neuroscience, University of Virginia, Charlottesville, Virginia, USA.
authors: J Julius
authors: JJ
authors: Zhu
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000502:physiology*; D051379:Mice; D009434:Neural Pathways / Q000502:physiology*; D009488:Neurosciences / Q000379:methods*; D062308:Optogenetics / Q000379:methods*; D018408:Patch-Clamp Techniques / Q000379:methods*; D013569:Synapses / Q000502:physiology*
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