Calcium Imaging of Microglial Network Activity in Stroke
| Autor: | Kathryn N. Kearns, M. Yashar S. Kalani, M. Filip Sluzewski, Scott T. Acton, Khadijeh A Sharifi, Petr Tvrdik |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Microglia Cell chemistry.chemical_element Calcium Biology Calcium in biology Green fluorescent protein 03 medical and health sciences 030104 developmental biology 0302 clinical medicine medicine.anatomical_structure Calcium imaging chemistry medicine Neuroscience 030217 neurology & neurosurgery Preclinical imaging Calcium signaling |
| Zdroj: | Microglia ISBN: 9781493996575 |
| DOI: | 10.1007/978-1-4939-9658-2_19 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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