Manseau Frédéric, Williams Sylvain
Department of Psychiatry, Douglas Mental Health University Institute, McGill University;
Department of Psychiatry, Douglas Mental Health University Institute, McGill University.
J Vis Exp. 2017 Aug 2(126):55851. doi: 10.3791/55851.
This protocol outlines the procedures for preparing and recording from the isolated whole hippocampus, of WT and transgenic mice, along with recent improvements in methodologies and applications for the study of theta oscillations. A simple characterization of the isolated hippocampal preparation is presented whereby the relationship between internal hippocampal theta oscillators is examined together with the activity of pyramidal cells, and GABAergic interneurons, of the cornu ammonis-1 (CA1) and subiculum (SUB) areas. Overall, we show that the isolated hippocampus is capable of generating intrinsic theta oscillations in vitro and that rhythmicity generated within the hippocampus can be precisely manipulated by optogenetic stimulation of parvalbumin-positive (PV) interneurons. The in vitro isolated hippocampal preparation offers a unique opportunity to use simultaneous field and intracellular patch-clamp recordings from visually-identified neurons to better understand the mechanisms underlying theta rhythm generation.
本方案概述了从野生型和转基因小鼠的离体完整海马体进行制备和记录的程序，以及近期在研究θ振荡的方法和应用方面的改进。文中给出了离体海马体制备的简单特征描述，据此研究了海马体内θ振荡器之间的关系，以及海马角-1（CA1）和下托（SUB）区域锥体细胞和GABA能中间神经元的活动。总体而言，我们表明离体海马体能够在体外产生内在的θ振荡，并且海马体内产生的节律性可以通过对小白蛋白阳性（PV）中间神经元的光遗传学刺激进行精确调控。体外离体海马体制备提供了一个独特的机会，可对视觉识别的神经元同时进行场电位和细胞内膜片钳记录，以更好地理解θ节律产生的机制。
