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CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections.

Publication ,  Journal Article
Cronan, MR; Rosenberg, AF; Oehlers, SH; Saelens, JW; Sisk, DM; Jurcic Smith, KL; Lee, S; Tobin, DM
Published in: Dis Model Mech
December 2015

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.

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Published In

Dis Model Mech

DOI

EISSN

1754-8411

Publication Date

December 2015

Volume

8

Issue

12

Start / End Page

1643 / 1650

Location

England

Related Subject Headings

  • Zebrafish
  • Tumor Necrosis Factor-alpha
  • Tuberculosis
  • Neovascularization, Physiologic
  • Mycobacterium tuberculosis
  • Mice, Inbred C57BL
  • Lung
  • Imaging, Three-Dimensional
  • Granuloma
  • Fluorescence
 

Citation

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Chicago
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MLA
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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. Dis Model Mech, 8(12), 1643–1650. https://doi.org/10.1242/dmm.021394
Cronan, Mark R., Allison F. Rosenberg, Stefan H. Oehlers, Joseph W. Saelens, Dana M. Sisk, Kristen L. Jurcic Smith, Sunhee Lee, and David M. Tobin. “CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections.Dis Model Mech 8, no. 12 (December 2015): 1643–50. 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. Dis Model Mech. 2015 Dec;8(12):1643–50.
Cronan, Mark R., et al. “CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections.Dis Model Mech, vol. 8, no. 12, Dec. 2015, pp. 1643–50. Pubmed, doi:10.1242/dmm.021394.
Cronan MR, Rosenberg AF, Oehlers SH, Saelens JW, Sisk DM, Jurcic Smith KL, Lee S, Tobin DM. CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections. Dis Model Mech. 2015 Dec;8(12):1643–1650.
Journal cover image

Published In

Dis Model Mech

DOI

EISSN

1754-8411

Publication Date

December 2015

Volume

8

Issue

12

Start / End Page

1643 / 1650

Location

England

Related Subject Headings

  • Zebrafish
  • Tumor Necrosis Factor-alpha
  • Tuberculosis
  • Neovascularization, Physiologic
  • Mycobacterium tuberculosis
  • Mice, Inbred C57BL
  • Lung
  • Imaging, Three-Dimensional
  • Granuloma
  • Fluorescence