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Elife. 2015 Jul 8;4:e07288. doi: 10.7554/eLife.07288.

Identification of polarized macrophage subsets in zebrafish.

Author information

1
Institut de Médecine Régénérative et Biothérapies, Institut national de la santé et de la recherche médicale, Montpellier, France.
2
Université de Montpellier, Montpellier, France.
3
Dynamique des Interactions Membranaires Normales et Pathologiques, Centre national de la recherche scientifique, Montpellier, France.
4
Macrophages et Développement de l'Immunité, Institut Pasteur, Paris, France.

Abstract

While the mammalian macrophage phenotypes have been intensively studied in vitro, the dynamic of their phenotypic polarization has never been investigated in live vertebrates. We used the zebrafish as a live model to identify and trail macrophage subtypes. We generated a transgenic line whose macrophages expressing tumour necrosis factor alpha (tnfa), a key feature of classically activated (M1) macrophages, express fluorescent proteins Tg(mpeg1:mCherryF/tnfa:eGFP-F). Using 4D-confocal microscopy, we showed that both aseptic wounding and Escherichia coli inoculation triggered macrophage recruitment, some of which started to express tnfa. RT-qPCR on Fluorescence Activated Cell Sorting (FACS)-sorted tnfa(+) and tnfa(-) macrophages showed that they, respectively, expressed M1 and alternatively activated (M2) mammalian markers. Fate tracing of tnfa(+) macrophages during the time-course of inflammation demonstrated that pro-inflammatory macrophages converted into M2-like phenotype during the resolution step. Our results reveal the diversity and plasticity of zebrafish macrophage subsets and underline the similarities with mammalian macrophages proposing a new system to study macrophage functional dynamic.

KEYWORDS:

developmental biology; immunology; live imaging; macrophages; stem cells; zebrafish

PMID:
26154973
PMCID:
PMC4521581
DOI:
10.7554/eLife.07288
[Indexed for MEDLINE]
Free PMC Article

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