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  Imaging Reporter Strategy to Monitor Gene Activation of Microglia Polarisation States under Stimulation

Collmann, F. M., Pijnenburg, R., Schneider, G., Schafer, C., Folz-Donahue, K., Kukat, C., et al. (2018). Imaging Reporter Strategy to Monitor Gene Activation of Microglia Polarisation States under Stimulation. J Neuroimmune Pharmacol, 13(3), 371-382. doi:10.1007/s11481-018-9789-2.

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 Creators:
Collmann, F. M.1, Author
Pijnenburg, R.1, Author
Schneider, G.1, Author
Schafer, C.1, Author
Folz-Donahue, K.1, Author
Kukat, C.1, Author
Hoehn, M.1, Author
Affiliations:
1Max Planck Institute for Biology of Ageing, Max Planck Society, Joseph-Stelzmann-Str. 9b, D-50931 Cologne, DE, ou_1942284              

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Free keywords: Bioluminescence imaging Fluorescence imaging Microglia Microglia polarisation Reporter imaging
 Abstract: Microglial cells as innate immune key players have a critical and unique role in neurodegenerative disorders. They strongly interact with their microenvironment in a complex manner and react to changes by switching their phenotype and functional activation states. In order to understand the development of brain diseases, it is imperative to elucidate up- or down-regulation of genes involved in microglia polarisation in time-profile by a simple-to-use strategy. Here, we present a new imaging strategy to follow promoter activity of genes involved in microglia polarisation. We lentivirally transduced BV-2 microglia cells in culture with constructs consisting of the induced nitric oxide synthase (iNOS), Fc gamma receptor III (Fcgr3) (both resembling the pro-inflammatory M1-like phenotype) or Chitinase-like 3 (Chil3/Ym1) (resembling the anti-inflammatory M2-like phenotype) promoters and stimulated transgenic cells with potent activators for pro- or anti-inflammatory response, such as lipopolysaccharide (LPS) + interferon gamma (IFN-gamma) or interleukin (IL)-4, respectively. Promoter activities upon polarisation phases were quantitatively assessed by the two imaging reporters Luc2 for bioluminescence and eGFP for fluorescence.

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 Dates: 2018-092018
 Publication Status: Issued
 Pages: -
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 Rev. Type: -
 Identifiers: Other: 29790106
DOI: 10.1007/s11481-018-9789-2
ISSN: 1557-1904 (Electronic)1557-1890 (Linking)
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Title: J Neuroimmune Pharmacol
Source Genre: Journal
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Pages: - Volume / Issue: 13 (3) Sequence Number: - Start / End Page: 371 - 382 Identifier: -