A new approach for ratiometric in vivo calcium imaging of microglia

Research output: Contribution to journalArticle

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.

Details

Authors
  • Bianca Brawek
  • Yajie Liang
  • Daria Savitska
  • Kaizhen Li
  • Natalie Fomin-Thunemann
  • Yury Kovalchuk
  • Elizabeta Zirdum
  • Johan Jakobsson
  • Olga Garaschuk
Organisations
External organisations
  • University of Tübingen
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Cell and Molecular Biology
  • Neurosciences
Original languageEnglish
Article number6030
JournalScientific Reports
Volume7
Issue number1
Publication statusPublished - 2017 Dec 1
Publication categoryResearch
Peer-reviewedYes