Calcium Imaging of Microglial Network Activity in Stroke

Methods in Molecular Biology
Petr TvrdikM Yashar S Kalani

Abstract

Calcium signaling plays a significant role in microglial activation. Genetically encoded calcium indicators (GECI) have been widely used for calcium imaging studies in many brain cell types, including neurons, astrocytes, and oligodendrocytes. However, microglial calcium imaging approaches have been hampered by idiosyncrasies of their gene expression and malleable cell properties. The generation of PC::G5-tdT, a Polr2a locus-based conditional mouse reporter of calcium, facilitated the deployment of GECI in microglia. When crossed with the Iba1(Aif1)-IRES-Cre line, all brain microglia of the progeny are labeled with the calcium indicator variant GCaMP5G and the red fluorescent protein tdTomato. This reporter system has enabled in vivo studies of intracellular calcium in large microglial cell populations in cerebral pathologies such as ischemic stroke. In this chapter, we outline specific guidelines for genetic, surgical, imaging, and data analysis aspects of microglial calcium monitoring of the ischemic cortex following middle cerebral artery occlusion.

Citations

Jan 8, 2020·Neurotherapeutics : the Journal of the American Society for Experimental NeuroTherapeutics·A P Coulibaly, J J Provencio
Nov 10, 2020·Stroke; a Journal of Cerebral Circulation·Lei LiuPetr Tvrdik
Oct 31, 2020·Frontiers in Neuroscience·Ricardo Augusto de Melo ReisFernando Garcia de Mello
May 30, 2021·Translational Stroke Research·Weijie ChenPeiying Li
Jun 10, 2021·Neuroscience Bulletin·Yang Zhang, Donghong Cui
Aug 19, 2021·Stroke; a Journal of Cerebral Circulation·Aminata P CoulibalyJ Javier Provencio

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