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Merrily they roll along


Neutrophils play a critical role in mediating injury following ischemia and reperfustion, but the dynamic regulation of leukocyte recruitment has been difficult to study in vivo. In this article, Li et al. used 2 photon microscopy to observe neutrophil trafficking in live mice following cardiac graft surgery. In these frames, neutrophils (yellow) are shown rolling along the endothelium of coronary veins. They also captured video of this movement.

Published June 19, 2012, by Kathryn Claiborn

Scientific Show Stopper

Related articles

Intravital 2-photon imaging of leukocyte trafficking in beating heart
Wenjun Li, … , Mark J. Miller, Daniel Kreisel
Wenjun Li, … , Mark J. Miller, Daniel Kreisel
Published July 2, 2012; First published June 18, 2012
Citation Information: J Clin Invest. 2012;122(7):2499-2508. https://doi.org/10.1172/JCI62970.
View: Text | PDF
Categories: Technical Advance Cardiology

Intravital 2-photon imaging of leukocyte trafficking in beating heart

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Abstract

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.

Authors

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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