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Recombinant WNTs differentially activate β-catenin-dependent and -independent signaling in mouse microglia-like cells.
MedLine Citation:
PMID:  21557822     Owner:  NLM     Status:  Publisher    
Aim:  The objective of this study is to compare the efficacy of different recombinant, commercially available WNTs with regard to WNT/β-catenin signaling, Dishevelled (DVL) and G protein activation and the induction of cell proliferation in a microglia-like cell line called N13. Method:  For detection of activated signaling molecules, cell lysates are analysed by immunoblotting. Further, we use 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 MTT 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 signaling in N13 cells. All WNTs induced the formation of phosphorylated and shifted 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. Conclusions:  WNTs show significant differences in their efficacy to activate β-catenin-dependent and -independent signaling. 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.
Michaela Bc Kilander; Carina Halleskog; Gunnar Schulte
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-5-9
Journal Detail:
Title:  Acta physiologica (Oxford, England)     Volume:  -     ISSN:  1748-1716     ISO Abbreviation:  -     Publication Date:  2011 May 
Date Detail:
Created Date:  2011-5-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101262545     Medline TA:  Acta Physiol (Oxf)     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2011 Scandinavian Physiological Society.
Karolinska Institutet, Dept Physiology and Pharmacology, Sec Receptor Biology & Signaling, S-17177 Stockholm, Sweden.
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