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Zebrafish Embryos as a Model to Study Bacterial Virulence

  • Jennifer Mesureur
  • Annette C. VergunstEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1197)

Abstract

In recent years the zebrafish has gained enormous attention in infection biology, and many protocols have been developed to study interaction of both human and fish pathogens, including viruses, fungi, and bacteria, with the host. Especially the extraordinary possibilities for live imaging of disease processes in the transparent embryos using fluorescent bacteria and cell-specific reporter fish combined with gene knockdown, transcriptome, and genetic studies have dramatically advanced our understanding of disease mechanisms. The zebrafish embryo is amenable to study virulence of both extracellular and facultative intracellular pathogens introduced through the technique of microinjection. Several protocols have been published that address the different sites of injection, antisense strategies, imaging, and production of transgenic fish in detail. Here we describe a protocol to study the virulence profiles, ranging from acute fatal to persistent, of bacteria belonging to the Burkholderia cepacia complex. This standard operating protocol combines simple survival assays, analysis of bacterial kinetics, analysis of the early innate immune response with qRT-PCR, and the use of transgenic reporter fish to study interactions with host phagocytes, and is also applicable to other pathogens.

Key words

Zebrafish Burkholderia cepacia complex Burkholderia cenocepacia Bacterial virulence Intracellular bacteria Infection profiles 

Notes

Acknowledgements

The authors would like to thank Nicolas Cubedo (INSERM, UMR_S710, Montpellier, France), Georges Lutfalla (CNRS, UMR5235 Montpellier, France), Annemarie Meijer, Erica Benard, and Michiel van der Vaart (Institute of Biology Leiden, The Netherlands) for helpful suggestions and discussion. U1047 was supported by the region Languedoc-Roussillon, INSERM, and the Université de Montpellier 1. AV was supported by grants from FRM, the region Languedoc-Roussillon (“Chercheur d’Avenir”), and the Marie-Curie Initial Training Network FishForPharma (PITN-GA-2011-289209). JM was supported by a grant from the French Ministry of Higher Education and Research.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.INSERM, U1047NîmesFrance
  2. 2.Université Montpellier 1, UFR MédecineNîmesFrance

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