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

England

2016-08-10T17:46:40Z

2015-12

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.

http://www.ncbi.nlm.nih.gov/pubmed/26449262

dmm.021394

1754-8411

https://hdl.handle.net/10161/12636

eng

The Company of Biologists

Dis Model Mech

10.1242/dmm.021394

CLARITY

Imaging

Infection

Mouse

Mycobacteria

PACT

Tuberculosis

Zebrafish

Aging

Animals

Blood Vessels

Disease Models, Animal

Female

Fluorescence

Granuloma

Imaging, Three-Dimensional

Lung

Mice, Inbred C57BL

Mycobacterium tuberculosis

Neovascularization, Physiologic

Tuberculosis

Tumor Necrosis Factor-alpha

Zebrafish

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

Journal article

Tobin, David M|0000-0003-3465-5518

http://www.ncbi.nlm.nih.gov/pubmed/26449262

1643

1650

12

Basic Science Departments

Clinical Science Departments

Duke

Duke Human Vaccine Institute

Immunology

Institutes and Centers

Molecular Genetics and Microbiology

Pathology

School of Medicine

Published

8