title: A high-throughput chemically induced inflammation assay in zebrafish.
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.
authors.affiliation: Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
authors.forename: Claudia A
authors.initials: CA
authors.lastname: d'Alençon
authors.forename: Oscar A
authors.initials: OA
authors.lastname: Peña
authors.forename: Christine
authors.initials: C
authors.lastname: Wittmann
authors.forename: Viviana E
authors.initials: VE
authors.lastname: Gallardo
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Jones
authors.forename: Felix
authors.initials: F
authors.lastname: Loosli
authors.forename: Urban
authors.initials: U
authors.lastname: Liebel
authors.forename: Clemens
authors.initials: C
authors.lastname: Grabher
authors.forename: Miguel L
authors.initials: ML
authors.lastname: Allende
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004625:Embryo, Nonmammalian; D057166:High-Throughput Screening Assays; D007155:Immunologic Factors / Q000009:adverse effects / Q000302:isolation & purification* / Q000494:pharmacology*; D007249:Inflammation / Q000139:chemically induced* / Q000276:immunology; D007962:Leukocytes / Q000502:physiology; D008954:Models, Biological; D020556:Neutrophil Infiltration / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D015027:Zebrafish / Q000276:immunology*
chemical_list: D000893:Anti-Inflammatory Agents; D007155:Immunologic Factors; D017382:Reactive Oxygen Species