A new approach for ratiometric in vivo calcium imaging of microglia

TitleA new approach for ratiometric in vivo calcium imaging of microglia
Publication TypeJournal Article
Year of Publication2017
AuthorsB, Brawek, Y Liang, D Savitska, K Li, N Fomin-Thunemann, Y Kovalchuk, E Zirdum, J Jakobsson, and O Garaschuk
JournalScientific Reports
Abstract

Microglia, resident immune cells of the brain, react to the presence of pathogens/danger signals with a large repertoire of functional responses including morphological changes, proliferation, chemotaxis, production/release of cytokines, and phagocytosis. In vitro studies suggest that many of these effector functions are Ca2+-dependent, but our knowledge about in vivo Ca2+ signalling in microglia is rudimentary. This is mostly due to technical reasons, as microglia largely resisted all attempts of in vivo labelling with Ca2+ indicators. Here, we introduce a novel approach, utilizing a microglia-specific microRNA-9-regulated viral vector, enabling the expression of a genetically-encoded ratiometric Ca2+ sensor Twitch-2B in microglia. The Twitch-2B-assisted in vivo imaging enables recording of spontaneous and evoked microglial Ca2+ signals and allows for the first time to monitor the steady state intracellular Ca2+ levels in microglia. Intact in vivo microglia show very homogenous and low steady state intracellular Ca2+ levels. However, the levels increase significantly after acute slice preparation and cell culturing along with an increase in the expression of activation markers CD68 and IL-1β. These data identify the steady state intracellular Ca2+ level as a versatile microglial activation marker, which is highly sensitive to the cell’s environment.

URLhttps://www.nature.com/articles/s41598-017-05952-3.epdf?author_access_token=c8wORfTR418fdhJv-5o8I9RgN0jAjWel9jnR3ZoTv0N7_DiSUU1QKjkBAgT-ilWfF9SSVZqdyrJ2AheRnhKOQGO05CEcrvS_1wW6rBEiS5JQ-t6pV2-YheV2T4lWc12hhU_-mSbXOjujGeWgLdWITA%3D%3D