A high-throughput chemically induced inflammation assay in zebrafish

Claudia A. d'Alençon, Oscar A. Peña, Christine Wittmann, Viviana E. Gallardo, Rebecca A. Jones, Felix Loosli, Urban Liebel*, Clemens Grabher, Miguel L. Allende

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

145 Citations (Scopus)

Abstract

Background: Studies on innate immunity have benefited from the introduction of zebrafish as a model system. Transgenic fish expressing fluorescent proteins in leukocyte populations allow direct, quantitative visualization of an inflammatory response in vivo. It has been proposed that this animal model can be used for high-throughput screens aimed at the identification of novel immunomodulatory lead compounds. However, current assays require invasive manipulation of fish individually, thus preventing high-content screening.Results: Here we show that specific, noninvasive damage to lateral line neuromast cells can induce a robust acute inflammatory response. Exposure of fish larvae to sublethal concentrations of copper sulfate selectively damages the sensory hair cell population inducing infiltration of leukocytes to neuromasts within 20 minutes. Inflammation can be assayed in real time using transgenic fish expressing fluorescent proteins in leukocytes or by histochemical assays in fixed larvae. We demonstrate the usefulness of this method for chemical and genetic screens to detect the effect of immunomodulatory compounds and mutations affecting the leukocyte response. Moreover, we transformed the assay into a high-throughput screening method by using a customized automated imaging and processing system that quantifies the magnitude of the inflammatory reaction.Conclusions: This approach allows rapid screening of thousands of compounds or mutagenized zebrafish for effects on inflammation and enables the identification of novel players in the regulation of innate immunity and potential lead compounds toward new immunomodulatory therapies. We have called this method the chemically induced inflammation assay, or ChIn assay.See Commentary article: http://www.biomedcentral.com/1741-7007/8/148.

Original languageEnglish
Article number151
JournalBMC Biology
Volume8
DOIs
Publication statusPublished - 22 Dec 2010

Bibliographical note

Funding Information:
We thank Catalina Lafourcade, Víctor Guzmán, Nadine Eschen and Sebastian Hoetzel for expert fish care, Florencio Espinoza for technical help and Luisa Pereiro for help with time-lapse imaging. Zebrafish strains were kindly provided by Stephen Renshaw, Phillip Crosier, John Rawls, Darren Gilmour, Paul Martin, Herwig Baier and Leonard Zon. This work was supported by grants to MA from Fondecyt (1070867), FONDAP (15090007), ICM (P06-039F), CORFO-Innova (09MCSS-6705), DFG-Conicyt 075-2009; to CD from UNAB (DI-01-09/1) and Fondecyt (24090004); to UL from Dopaminet (EU FP7 223744); and to CG by a Marie Curie International Reintegration Grant (EU FP7; PIRG07-GA-2010-267552).

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