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成年小鼠齿状回颗粒细胞兴奋性的昼夜节律变化的电生理测量方案。

Protocol for electrophysiological measurements of circadian changes in excitability in dentate granule cells from adult mice.

机构信息

Department of Neurobiology and McKnight Institute, University of Alabama at Birmingham, Birmingham, AL 35294, USA.

Department of Neurobiology and McKnight Institute, University of Alabama at Birmingham, Birmingham, AL 35294, USA.

出版信息

STAR Protoc. 2024 Sep 20;5(3):103255. doi: 10.1016/j.xpro.2024.103255. Epub 2024 Aug 14.

Abstract

Many types of neurons exhibit a daily rhythm of intrinsic excitability. Here, we present a protocol for assessing circadian regulation of dentate granule cell excitability using a mouse model for conditional knockout of the molecular clock protein BMAL1. We describe steps for obtaining healthy oblique horizontal slices that contain the hippocampus and measuring intrinsic excitability and synaptic potentials by combining whole-cell patch-clamp recordings and perforant-path electric stimulation. We then detail procedures for validating single-cell genetic deletion of Bmal1 by immunohistochemistry. For complete details on the use and execution of this protocol, please refer to Gonzalez et al..

摘要

许多类型的神经元表现出内在兴奋性的日常节律。在这里,我们提出了一种使用条件敲除分子钟蛋白 BMAL1 的小鼠模型来评估齿状回颗粒细胞兴奋性的生物钟调节的方案。我们描述了获得包含海马体的健康斜水平切片的步骤,并通过结合全细胞膜片钳记录和穿通路径电刺激来测量内在兴奋性和突触电位。然后,我们详细介绍了通过免疫组织化学验证 Bmal1 单细胞基因缺失的程序。有关此方案的使用和执行的完整详细信息,请参阅 Gonzalez 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8d57/11367413/f24903e80b49/fx1.jpg

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