A new approach for ratiometric in vivo calcium imaging of microglia
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...
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Nature Portfolio
2017
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oai:doaj.org-article:d571013a91024fb297deb6a6ac58ad7b2021-12-02T15:06:04ZA new approach for ratiometric in vivo calcium imaging of microglia10.1038/s41598-017-05952-32045-2322https://doaj.org/article/d571013a91024fb297deb6a6ac58ad7b2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05952-3https://doaj.org/toc/2045-2322Abstract 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.Bianca BrawekYajie LiangDaria SavitskaKaizhen LiNatalie Fomin-ThunemannYury KovalchukElizabeta ZirdumJohan JakobssonOlga GaraschukNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017) |
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DOAJ |
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Medicine R Science Q |
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Medicine R Science Q Bianca Brawek Yajie Liang Daria Savitska Kaizhen Li Natalie Fomin-Thunemann Yury Kovalchuk Elizabeta Zirdum Johan Jakobsson Olga Garaschuk A new approach for ratiometric in vivo calcium imaging of microglia |
| description |
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. |
| format |
article |
| author |
Bianca Brawek Yajie Liang Daria Savitska Kaizhen Li Natalie Fomin-Thunemann Yury Kovalchuk Elizabeta Zirdum Johan Jakobsson Olga Garaschuk |
| author_facet |
Bianca Brawek Yajie Liang Daria Savitska Kaizhen Li Natalie Fomin-Thunemann Yury Kovalchuk Elizabeta Zirdum Johan Jakobsson Olga Garaschuk |
| author_sort |
Bianca Brawek |
| title |
A new approach for ratiometric in vivo calcium imaging of microglia |
| title_short |
A new approach for ratiometric in vivo calcium imaging of microglia |
| title_full |
A new approach for ratiometric in vivo calcium imaging of microglia |
| title_fullStr |
A new approach for ratiometric in vivo calcium imaging of microglia |
| title_full_unstemmed |
A new approach for ratiometric in vivo calcium imaging of microglia |
| title_sort |
new approach for ratiometric in vivo calcium imaging of microglia |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/d571013a91024fb297deb6a6ac58ad7b |
| work_keys_str_mv |
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| _version_ |
1718388612823777280 |