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Recombinant WNTs differentially activate β‐catenin‐dependent and ‐independent signalling in mouse microglia‐like cells
Author(s) -
Kilander M. B. C.,
Halleskog C.,
Schulte G.
Publication year - 2011
Publication title -
acta physiologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.591
H-Index - 116
eISSN - 1748-1716
pISSN - 1748-1708
DOI - 10.1111/j.1748-1716.2011.02324.x
Subject(s) - wnt signaling pathway , microbiology and biotechnology , microglia , heterotrimeric g protein , catenin , neuroinflammation , biology , dishevelled , beta catenin , signal transduction , frizzled , g protein , immunology , inflammation
Abstract Aim: The objective of this study was to compare the efficacy of different recombinant, commercially available Wingless/Int‐1 (WNTs) with regard to WNT/β‐catenin signalling, dishevelled (DVL) and G protein activation and the induction of cell proliferation in a microglia‐like cell line called N13. Methods: For detection of activated signalling molecules, cell lysates are analysed by immunoblotting. Furthermore, we used a [γ 35 S] GTP binding assay to monitor the exchange of GDP for GTP in heterotrimeric G proteins in N13 membrane preparations. Cell proliferation was assessed by the 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay measuring mitochondrial function, which is proportional to the amount of viable cells. Results: Of the WNTs tested (WNT‐3A, ‐4, ‐5A, ‐5B, ‐7A,‐9B), only WNT‐3A activated WNT/β‐catenin signalling in N13 cells. All WNTs induced the formation of P hosphorylated and S hifted DVL (PS‐DVL) and the activation of heterotrimeric G proteins with variable efficacies. WNT‐5A and WNT‐9B, which had the highest efficacy in the G protein assay, also induced N13 cell proliferation. Conclusion: WNTs show significant differences in their efficacy to activate β‐catenin‐dependent and ‐independent signalling. The WNTs tested are present during maturation of the central nervous system and/or in the adult brain and are thus potential regulators of microglia‐mediated neuroinflammation.