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Wnt3a induces exosome secretion from primary cultured rat microglia

BACKGROUND: Microglia, the immune effector cells of the CNS and the signaling molecule Wnt, both play critical roles in neurodevelopment and neurological disease. Here we describe the inducible release of exosomes from primary cultured rat microglia following treatment with recombinant carrier-free...

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Autores principales: Hooper, Claudie, Sainz-Fuertes, Ricardo, Lynham, Steven, Hye, Abdul, Killick, Richard, Warley, Alice, Bolondi, Cecilia, Pocock, Jennifer, Lovestone, Simon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3541220/
https://www.ncbi.nlm.nih.gov/pubmed/23173708
http://dx.doi.org/10.1186/1471-2202-13-144
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author Hooper, Claudie
Sainz-Fuertes, Ricardo
Lynham, Steven
Hye, Abdul
Killick, Richard
Warley, Alice
Bolondi, Cecilia
Pocock, Jennifer
Lovestone, Simon
author_facet Hooper, Claudie
Sainz-Fuertes, Ricardo
Lynham, Steven
Hye, Abdul
Killick, Richard
Warley, Alice
Bolondi, Cecilia
Pocock, Jennifer
Lovestone, Simon
author_sort Hooper, Claudie
collection PubMed
description BACKGROUND: Microglia, the immune effector cells of the CNS and the signaling molecule Wnt, both play critical roles in neurodevelopment and neurological disease. Here we describe the inducible release of exosomes from primary cultured rat microglia following treatment with recombinant carrier-free Wnt3a. RESULTS: Wnt3a was internalised into microglia, being detectable in early endosomes, and secreted in exosomes through a GSK3-independent mechanism. Electron microscopy demonstrated that exosomes were elliptical, electron-dense (100 nm) vesicles that coalesced with time in vitro. In contrast to microglia, primary cortical neurons released exosomes constitutively and the quantity of exosomes released was not altered by Wnt3a treatment. The proteomic profile of the microglial-derived exosomes was characterised using liquid chromatography-tandem mass spectrometry (LC/MS/MS) and the vesicles were found to be associated with proteins involved in cellular architecture, metabolism, protein synthesis and protein degradation including β-actin, glyceraldehyde-3-phosphate dehydrogenase, ribosomal subunits and ubiquitin (45 proteins in total). Unlike lipopolysaccharide, Wnt3a did not induce a neurotoxic, pro-inflammatory phenotype in primary microglia. CONCLUSION: These findings reveal a novel mechanism through which Wnt3a signals in microglia resulting in the release of exosomes loaded with proteinaceous cargo.
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spelling pubmed-35412202013-01-11 Wnt3a induces exosome secretion from primary cultured rat microglia Hooper, Claudie Sainz-Fuertes, Ricardo Lynham, Steven Hye, Abdul Killick, Richard Warley, Alice Bolondi, Cecilia Pocock, Jennifer Lovestone, Simon BMC Neurosci Research Article BACKGROUND: Microglia, the immune effector cells of the CNS and the signaling molecule Wnt, both play critical roles in neurodevelopment and neurological disease. Here we describe the inducible release of exosomes from primary cultured rat microglia following treatment with recombinant carrier-free Wnt3a. RESULTS: Wnt3a was internalised into microglia, being detectable in early endosomes, and secreted in exosomes through a GSK3-independent mechanism. Electron microscopy demonstrated that exosomes were elliptical, electron-dense (100 nm) vesicles that coalesced with time in vitro. In contrast to microglia, primary cortical neurons released exosomes constitutively and the quantity of exosomes released was not altered by Wnt3a treatment. The proteomic profile of the microglial-derived exosomes was characterised using liquid chromatography-tandem mass spectrometry (LC/MS/MS) and the vesicles were found to be associated with proteins involved in cellular architecture, metabolism, protein synthesis and protein degradation including β-actin, glyceraldehyde-3-phosphate dehydrogenase, ribosomal subunits and ubiquitin (45 proteins in total). Unlike lipopolysaccharide, Wnt3a did not induce a neurotoxic, pro-inflammatory phenotype in primary microglia. CONCLUSION: These findings reveal a novel mechanism through which Wnt3a signals in microglia resulting in the release of exosomes loaded with proteinaceous cargo. BioMed Central 2012-11-23 /pmc/articles/PMC3541220/ /pubmed/23173708 http://dx.doi.org/10.1186/1471-2202-13-144 Text en Copyright ©2012 Hooper et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Hooper, Claudie
Sainz-Fuertes, Ricardo
Lynham, Steven
Hye, Abdul
Killick, Richard
Warley, Alice
Bolondi, Cecilia
Pocock, Jennifer
Lovestone, Simon
Wnt3a induces exosome secretion from primary cultured rat microglia
title Wnt3a induces exosome secretion from primary cultured rat microglia
title_full Wnt3a induces exosome secretion from primary cultured rat microglia
title_fullStr Wnt3a induces exosome secretion from primary cultured rat microglia
title_full_unstemmed Wnt3a induces exosome secretion from primary cultured rat microglia
title_short Wnt3a induces exosome secretion from primary cultured rat microglia
title_sort wnt3a induces exosome secretion from primary cultured rat microglia
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3541220/
https://www.ncbi.nlm.nih.gov/pubmed/23173708
http://dx.doi.org/10.1186/1471-2202-13-144
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