ZFIN ID: ZDB-PUB-160821-3
Infection of zebrafish embryos with live fluorescent Streptococcus pneumoniae as a real-time pneumococcal meningitis model
Jim, K.K., Engelen-Lee, J., van der Sar, A.M., Bitter, W., Brouwer, M.C., van der Ende, A., Veening, J.W., van de Beek, D., Vandenbroucke-Grauls, C.M.
Date: 2016
Source: Journal of neuroinflammation   13: 188 (Journal)
Registered Authors: Bitter, Wilbert, van der Sar, Astrid M.
Keywords: Host-microbe interaction, Live cell imaging, Pneumococcal meningitis, Pneumolysin, Streptococcus pneumoniae, Zebrafish
MeSH Terms:
  • Age Factors
  • Animals
  • Animals, Genetically Modified
  • Blood-Brain Barrier/microbiology
  • Blood-Brain Barrier/pathology
  • Disease Models, Animal
  • Embryo, Nonmammalian/microbiology
  • Gene Expression Regulation, Developmental/genetics*
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism
  • Immunity, Innate/physiology*
  • Luminescent Proteins/genetics
  • Luminescent Proteins/metabolism
  • Meningitis, Pneumococcal/genetics
  • Meningitis, Pneumococcal/microbiology*
  • Meningitis, Pneumococcal/mortality
  • Meningitis, Pneumococcal/pathology*
  • Streptococcus pneumoniae/pathogenicity*
  • Zebrafish
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed: 27542968 Full text @ J Neuroinflammation
Streptococcus pneumoniae is one of the most important causes of bacterial meningitis, an infection where unfavourable outcome is driven by bacterial and host-derived toxins. In this study, we developed and characterized a pneumococcal meningitis model in zebrafish embryos that allows for real-time investigation of early host-microbe interaction.
Zebrafish embryos were infected in the caudal vein or hindbrain ventricle with green fluorescent wild-type S. pneumoniae D39 or a pneumolysin-deficient mutant. The kdrl:mCherry transgenic zebrafish line was used to visualize the blood vessels, whereas phagocytic cells were visualized by staining with far red anti-L-plastin or in mpx:GFP/mpeg1:mCherry zebrafish, that have green fluorescent neutrophils and red fluorescent macrophages. Imaging was performed by fluorescence confocal and time-lapse microscopy.
After infection by caudal vein, we saw focal clogging of the pneumococci in the blood vessels and migration of bacteria through the blood-brain barrier into the subarachnoid space and brain tissue. Infection with pneumolysin-deficient S. pneumoniae in the hindbrain ventricle showed attenuated growth and migration through the brain as compared to the wild-type strain. Time-lapse and confocal imaging revealed that the initial innate immune response to S. pneumoniae in the subarachnoid space mainly consisted of neutrophils and that pneumolysin-mediated cytolytic activity caused a marked reduction of phagocytes.
This new meningitis model permits detailed analysis and visualization of host-microbe interaction in pneumococcal meningitis in real time and is a very promising tool to further our insights in the pathogenesis of pneumococcal meningitis.