使用基因和病毒方法在小鼠大脑中可视化编码不同事件的三个不同神经元群体的方案。

Protocol to visualize three distinct neuronal ensembles encoding different events in the mouse brain using genetic and viral approaches.

作者信息

Lesuis Sylvie L, Rashid Asim J, Hoorn Annelies, Park Sungmo, Mocle Andrew J, Torelli Angelica M, DeCristofaro Antonietta, Frankland Paul W, Hill Matthew N, Josselyn Sheena A

机构信息

Program in Neurosciences & Mental Health, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada; Department of Cellular and Computational Neuroscience, Swammerdam Institute for Life Sciences, Amsterdam Neuroscience, University of Amsterdam, Amsterdam 1090 GE, the Netherlands.

Program in Neurosciences & Mental Health, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada.

出版信息

STAR Protoc. 2025 Apr 8;6(2):103747. doi: 10.1016/j.xpro.2025.103747.

DOI:10.1016/j.xpro.2025.103747

PMID:40202842

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12008573/

Abstract

Activity tagging of neuronal ensembles has become an important tool in neuroscience. Here, we present a protocol for visualizing separate neuronal ensembles active during three distinct phases of a memory in transgenic mice. We describe steps to label active neurons using viral microinjection, inducing GFP expression under the robust activity marker (RAM) promoter, and transgenic mice, inducing tdTomato (TdT) expression, and immunohistochemical (IHC) visualization of endogenous cFos expression. We then detail procedures for preparation of tissue, imaging, and quantification of memory events. For complete details on the use and execution of this protocol, please refer to Lesuis et al..

摘要

神经元集群的活动标记已成为神经科学中的一项重要工具。在此，我们展示了一种用于可视化转基因小鼠在记忆的三个不同阶段活跃的不同神经元集群的方法。我们描述了使用病毒显微注射标记活跃神经元的步骤，在强活动标记（RAM）启动子下诱导绿色荧光蛋白（GFP）表达，以及使用转基因小鼠诱导tdTomato（TdT）表达，并通过免疫组织化学（IHC）可视化内源性cFos表达。然后，我们详细介绍了组织制备、成像和记忆事件定量的程序。有关此方法的使用和执行的完整详细信息，请参考勒叙伊等人的研究。