PUBLICATION

Modeling Tuberculosis in Mycobacterium marinum Infected Adult Zebrafish

Authors
Luukinen, H., Hammarén, M.M., Vanha-Aho, L.M., Parikka, M.
ID
ZDB-PUB-181023-7
Date
2018
Source
Journal of visualized experiments : JoVE   (140): (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • Animals
  • Disease Models, Animal*
  • Gene Expression
  • Host-Pathogen Interactions
  • Mycobacterium Infections, Nontuberculous/genetics
  • Mycobacterium Infections, Nontuberculous/immunology*
  • Mycobacterium Infections, Nontuberculous/microbiology*
  • Mycobacterium marinum/genetics
  • Mycobacterium marinum/physiology*
  • Real-Time Polymerase Chain Reaction
  • Zebrafish*
PubMed
30346391 Full text @ J. Vis. Exp.
Abstract
Mycobacterium tuberculosis is currently the deadliest human pathogen causing 1.7 million deaths and 10.4 million infections every year. Exposure to this bacterium causes a wide disease spectrum in humans ranging from a sterilized infection to an actively progressing deadly disease. The most common form is the latent tuberculosis, which is asymptomatic, but has the potential to reactivate into a fulminant disease. Adult zebrafish and its natural pathogen Mycobacterium marinum have recently proven to be an applicable model to study the wide disease spectrum of tuberculosis. Importantly, spontaneous latency and reactivation as well as adaptive immune responses in the context of mycobacterial infection can be studied in this model. In this article, we describe methods for the experimental infection of adult zebrafish, the collection of internal organs for the extraction of nucleic acids for the measurement of mycobacterial loads and host immune responses by quantitative PCR. The in-house-developed, M. marinum-specific qPCR assay is more sensitive than the traditional plating methods as it also detects DNA from non-dividing, dormant or recently dead mycobacteria. As both DNA and RNA are extracted from the same individual, it is possible to study the relationships between the diseased state, and the host and pathogen gene-expression. The adult zebrafish model for tuberculosis thus presents itself as a highly applicable, non-mammalian in vivo system to study host-pathogen interactions.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping