Document Detail

Intravital 2-photon imaging of leukocyte trafficking in beating heart.
MedLine Citation:
PMID:  22706307     Owner:  NLM     Status:  MEDLINE    
Two-photon intravital microscopy has substantially broadened our understanding of tissue- and organ-specific differences in the regulation of inflammatory responses. However, little is known about the dynamic regulation of leukocyte recruitment into inflamed heart tissue, largely due to technical difficulties inherent in imaging moving tissue. Here, we report a method for imaging beating murine hearts using intravital 2-photon microscopy. Using this method, we visualized neutrophil trafficking at baseline and during inflammation. Ischemia reperfusion injury induced by transplantation or transient coronary artery ligation led to recruitment of neutrophils to the heart, their extravasation from coronary veins, and infiltration of the myocardium where they formed large clusters. Grafting hearts containing mutant ICAM-1, a ligand important for neutrophil recruitment, reduced the crawling velocities of neutrophils within vessels, and markedly inhibited their extravasation. Similar impairment was seen with the inhibition of Mac-1, a receptor for ICAM-1. Blockade of LFA-1, another ICAM-1 receptor, prevented neutrophil adherence to endothelium and extravasation in heart grafts. As inflammatory responses in the heart are of great relevance to public health, this imaging approach holds promise for studying cardiac-specific mechanisms of leukocyte recruitment and identifying novel therapeutic targets for treating heart disease.
Wenjun Li; Ruben G Nava; Alejandro C Bribriesco; Bernd H Zinselmeyer; Jessica H Spahn; Andrew E Gelman; Alexander S Krupnick; Mark J Miller; Daniel Kreisel
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-06-18
Journal Detail:
Title:  The Journal of clinical investigation     Volume:  122     ISSN:  1558-8238     ISO Abbreviation:  J. Clin. Invest.     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-07-03     Completed Date:  2012-09-11     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  7802877     Medline TA:  J Clin Invest     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2499-508     Citation Subset:  AIM; IM    
Department of Surgery, Washington University School of Medicine, St. Louis, MO, USA.
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MeSH Terms
Cardiac Imaging Techniques / methods
Coronary Vessels / cytology
Green Fluorescent Proteins / biosynthesis
Heart Transplantation / immunology,  physiology
Leukocyte Rolling
Mice, Inbred C57BL
Microscopy, Fluorescence, Multiphoton / methods
Myocardial Contraction
Myocardium / cytology*,  immunology
Neutrophil Infiltration*
Quantum Dots
Recombinant Proteins / biosynthesis
Time-Lapse Imaging
Grant Support
Reg. No./Substance:
0/Recombinant Proteins; 147336-22-9/Green Fluorescent Proteins

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