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

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 developm...

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Detalles Bibliográficos
Autores principales: Collmann, Franziska M., Pijnenburg, Rory, Schneider, Gabriele, Schäfer, Cordula, Folz-Donahue, Kat, Kukat, Christian, Hoehn, Mathias
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6096558/
https://www.ncbi.nlm.nih.gov/pubmed/29790106
http://dx.doi.org/10.1007/s11481-018-9789-2
Descripción
Sumario: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-γ) 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11481-018-9789-2) contains supplementary material, which is available to authorized users.