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

dc.contributor.author Cronan, MR
dc.contributor.author Jurcic Smith, KL
dc.contributor.author Lee, S
dc.contributor.author Oehlers, SH
dc.contributor.author Rosenberg, AF
dc.contributor.author Saelens, Joseph W
dc.contributor.author Sisk, DM
dc.contributor.author Tobin, DM
dc.coverage.spatial England
dc.date.accessioned 2016-08-10T17:46:40Z
dc.date.issued 2015-12
dc.identifier http://www.ncbi.nlm.nih.gov/pubmed/26449262
dc.identifier dmm.021394
dc.identifier.uri https://hdl.handle.net/10161/12636
dc.description.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.
dc.language eng
dc.relation.ispartof Dis Model Mech
dc.relation.isversionof 10.1242/dmm.021394
dc.subject CLARITY
dc.subject Imaging
dc.subject Infection
dc.subject Mouse
dc.subject Mycobacteria
dc.subject PACT
dc.subject Tuberculosis
dc.subject Zebrafish
dc.subject Aging
dc.subject Animals
dc.subject Blood Vessels
dc.subject Disease Models, Animal
dc.subject Female
dc.subject Fluorescence
dc.subject Granuloma
dc.subject Imaging, Three-Dimensional
dc.subject Lung
dc.subject Mice, Inbred C57BL
dc.subject Mycobacterium tuberculosis
dc.subject Neovascularization, Physiologic
dc.subject Tuberculosis
dc.subject Tumor Necrosis Factor-alpha
dc.subject Zebrafish
dc.title CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections.
dc.type Journal article
pubs.author-url http://www.ncbi.nlm.nih.gov/pubmed/26449262
pubs.begin-page 1643
pubs.end-page 1650
pubs.issue 12
pubs.organisational-group Basic Science Departments
pubs.organisational-group Clinical Science Departments
pubs.organisational-group Duke
pubs.organisational-group Duke Human Vaccine Institute
pubs.organisational-group Immunology
pubs.organisational-group Institutes and Centers
pubs.organisational-group Molecular Genetics and Microbiology
pubs.organisational-group Pathology
pubs.organisational-group School of Medicine
pubs.publication-status Published
pubs.volume 8
dc.identifier.eissn 1754-8411


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