CLarity and pact-based imaging of adult zebrafish and mouse for whole-Animal analysis of infections

Mark R. Cronan, Allison F. Rosenberg, Stefan H. Oehlers, Joseph W. Saelens, Dana M. Sisk, Kristen L. Jurcic Smith, Sunhee Lee, David M. Tobin

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Visualization of infection and the associated host response has been challenging in adult vertebrates. Owing to their transparency, zebrafish larvae have been used to directly observe infection in vivo; however, such larvae have not yet developed a functional adaptive immune system. Cells involved in adaptive immunity mature later and have therefore been difficult to access optically in intact animals. Thus, the study of many aspects of vertebrate infection requires dissection of adult organs or ex vivo isolation of immune cells. Recently, CLARITY and PACT (passive clarity technique) methodologies have enabled clearing and direct visualization of dissected organs. Here, we show that these techniques can be applied to image host-pathogen interactions directly in whole animals. CLARITY and PACT-based clearing of whole adult zebrafish and Mycobacterium tuberculosis-infected mouse lungs enables imaging of mycobacterial granulomas deep within tissue to a depth of more than 1 mm. Using established transgenic lines, we were able to image normal and pathogenic structures and their surrounding host context at high resolution. We identified the three-dimensional organization of granuloma-associated angiogenesis, an important feature of mycobacterial infection, and characterized the induction of the cytokine tumor necrosis factor (TNF) within the granuloma using an established fluorescent reporter line. We observed heterogeneity in TNF induction within granuloma macrophages, consistent with an evolving view of the tuberculous granuloma as a non-uniform, heterogeneous structure. Broad application of this technique will enable new understanding of host-pathogen interactions in situ.

Original languageEnglish (US)
Pages (from-to)1643-1650
Number of pages8
JournalDMM Disease Models and Mechanisms
Volume8
Issue number12
DOIs
StatePublished - Dec 1 2015
Externally publishedYes

Fingerprint

Pathogens
Zebrafish
Granuloma
Animals
Visualization
Tumor Necrosis Factor-alpha
Imaging techniques
Dissection
Macrophages
Immune system
Host-Pathogen Interactions
Infection
Transparency
Larva
Vertebrates
Tissue
Cytokines
Cell Separation
Adaptive Immunity
Mycobacterium tuberculosis

Keywords

  • Clarity
  • Imaging
  • Infection
  • Mouse
  • Mycobacteria
  • PACT
  • Tuberculosis
  • Zebrafish

ASJC Scopus subject areas

  • Neuroscience (miscellaneous)
  • Medicine (miscellaneous)
  • Immunology and Microbiology (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Cronan, M. R., Rosenberg, A. F., Oehlers, S. H., Saelens, J. W., Sisk, D. M., Jurcic Smith, K. L., ... Tobin, D. M. (2015). CLarity and pact-based imaging of adult zebrafish and mouse for whole-Animal analysis of infections. DMM Disease Models and Mechanisms, 8(12), 1643-1650. https://doi.org/10.1242/dmm.021394

CLarity and pact-based imaging of adult zebrafish and mouse for whole-Animal analysis of infections. / Cronan, Mark R.; Rosenberg, Allison F.; Oehlers, Stefan H.; Saelens, Joseph W.; Sisk, Dana M.; Jurcic Smith, Kristen L.; Lee, Sunhee; Tobin, David M.

In: DMM Disease Models and Mechanisms, Vol. 8, No. 12, 01.12.2015, p. 1643-1650.

Research output: Contribution to journalArticle

Cronan, MR, Rosenberg, AF, Oehlers, SH, Saelens, JW, Sisk, DM, Jurcic Smith, KL, Lee, S & Tobin, DM 2015, 'CLarity and pact-based imaging of adult zebrafish and mouse for whole-Animal analysis of infections', DMM Disease Models and Mechanisms, vol. 8, no. 12, pp. 1643-1650. https://doi.org/10.1242/dmm.021394
Cronan MR, Rosenberg AF, Oehlers SH, Saelens JW, Sisk DM, Jurcic Smith KL et al. CLarity and pact-based imaging of adult zebrafish and mouse for whole-Animal analysis of infections. DMM Disease Models and Mechanisms. 2015 Dec 1;8(12):1643-1650. https://doi.org/10.1242/dmm.021394
Cronan, Mark R. ; Rosenberg, Allison F. ; Oehlers, Stefan H. ; Saelens, Joseph W. ; Sisk, Dana M. ; Jurcic Smith, Kristen L. ; Lee, Sunhee ; Tobin, David M. / CLarity and pact-based imaging of adult zebrafish and mouse for whole-Animal analysis of infections. In: DMM Disease Models and Mechanisms. 2015 ; Vol. 8, No. 12. pp. 1643-1650.
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