Skip to Main content Skip to Navigation
Journal articles

Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos

Abstract : Zebrafish (Danio rerio) embryos are increasingly used as an infection model to study the function of the vertebrate innate immune system in host-pathogen interactions. The ease of obtaining large numbers of embryos, their accessibility due to external development, their optical transparency as well as the availability of a wide panoply of genetic/immunological tools and transgenic reporter line collections, contribute to the versatility of this model. In this respect, the present manuscript describes the use of zebrafish as an in vivo model system to investigate the chronology of Mycobacterium abscessus infection. This human pathogen can exist either as smooth (S) or rough (R) variants, depending on cell wall composition, and their respective virulence can be imaged and compared in zebrafish embryos and larvae. Micro-injection of either S or R fluorescent variants directly in the blood circulation via the caudal vein, leads to chronic or acute/lethal infections, respectively. This biological system allows high resolution visualization and analysis of the role of mycobacterial cording in promoting abscess formation. In addition, the use of fluorescent bacteria along with transgenic zebrafish lines harbouring fluorescent macrophages produces a unique opportunity for multi-color imaging of the host-pathogen interactions. This article describes detailed protocols for the preparation of homogenous M. abscessus inoculum and for intravenous injection of zebrafish embryos for subsequent fluorescence imaging of the interaction with macrophages. These techniques open the avenue to future investigations involving mutants defective in cord formation and are dedicated to understand how this impacts on M. abscessus pathogenicity in a whole vertebrate.
Complete list of metadata

Cited literature [43 references]  Display  Hide  Download

https://hal.umontpellier.fr/hal-02086982
Contributor : Georges Lutfalla Connect in order to contact the contributor
Submitted on : Monday, April 1, 2019 - 4:49:17 PM
Last modification on : Thursday, October 14, 2021 - 12:04:02 PM
Long-term archiving on: : Tuesday, July 2, 2019 - 5:50:06 PM

File

bernutJOVE2015.pdf
Publisher files allowed on an open archive

Identifiers

Citation

Audrey Bernut, Christian Dupont, Alain Sahuquet, Jean-Louis Herrmann, Georges Lutfalla, et al.. Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos. Journal of visualized experiments : JoVE, JoVE, 2015, pp.e53130. ⟨10.3791/53130⟩. ⟨hal-02086982⟩

Share

Metrics

Record views

414

Files downloads

1125