Dynamics of neutrophil extravasation and vascular permeability are uncoupled during aseptic cutaneous wounding.
Transport of macromolecules and transmigration of leukocytes across vascular endothelium are regulated by a tight molecular junction, but the mechanisms by which these two inflammatory events are differentially controlled in time and magnitude during aseptic cutaneous wounding remain elusive. A real-time fluorescence imaging technique was developed to simultaneously track influx of Alexa 680-labeled albumin and genetically tagged enhanced green fluorescent protein-neutrophils [polymorphonuclear neutrophils (PMN)] within the wound bed. Vascular permeability increased approximately threefold more rapidly than the rate of PMN influx, reaching a maximum at 12 h, on the order of approximately 0.15% per minute versus approximately 0.05% per minute for PMN influx, which peaked at 18 h. Systemic depletion of PMN with antibody blocked their extravasation to the wound but did not alter the increase in vascular permeability. In contrast, pretreatment with antiplatelet GPIb decreased permeability by 25% and PMN influx by 50%. Hyperpermeability stimulated by the endothelium-specific agonists VEGF or thrombin at 24 h postwounding was completely inhibited by blocking Rho-kinase-dependent signaling, whereas less inhibition was observed at 1 h and neutrophil influx was not perturbed. These data suggest that in aseptic wounds, the endothelium maintains a tight junctional barrier to protein leakage that is independent of neutrophil transmigration, partially dependent on circulating platelets, and associated with Rho-kinase-dependent signaling.
Kim
Min-Ho
MH
Dept. of Biomedical Engineering, Univ. of California at Davis, 451 E. Health Sciences Dr., Davis, CA 95616, USA.
Curry
Fitz-Roy E
FR
Simon
Scott I
SI
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism; D002199:Capillary Permeability; D062109:Dermatologic Surgical Procedures; D004195:Disease Models, Animal; D004730:Endothelium, Vascular / Q000276:immunology / Q000378:metabolism / Q000503:physiopathology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D007365:Intercellular Junctions / Q000378:metabolism; D036904:Leukocyte Rolling; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism; D015539:Platelet Activation; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D015398:Signal Transduction; D012867:Skin / Q000098:blood supply* / Q000503:physiopathology*; D013917:Thrombin / Q000378:metabolism; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D014945:Wound Healing; D054460:rho-Associated Kinases / Q000378:metabolism
D019038:Platelet Glycoprotein GPIb-IX Complex; D042461:Vascular Endothelial Growth Factor A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D012710:Serum Albumin, Bovine; D054460:rho-Associated Kinases; D013917:Thrombin