PDF(Pubmed)
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.1.
摘要：
许多类型的神经元表现出内在兴奋性的每日节律。这里,我们提出了一种使用小鼠模型评估齿状颗粒细胞兴奋性的昼夜节律调节的方案，该模型用于条件敲除分子时钟蛋白BMAL1。我们描述了通过结合全细胞膜片钳记录和穿孔路径电刺激来获得包含海马的健康倾斜水平切片并测量内在兴奋性和突触电位的步骤。然后，我们详细介绍了通过免疫组织化学验证Bmal1单细胞遗传缺失的程序。有关此协议的使用和执行的完整详细信息，PleaserefertoGonzalezetal.1.
