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Articles in PresS, published online ahead of print August 8, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00787.2001
Submitted on September 4, 2001
Accepted on July 25, 2002
1 Department of Human Physiology, University of California Davis, Davis, CA, USA
2 Department of Laboratory Medicine, University of California San Francisco, San Francisco, CA, USA
3 Department of Physiology and Pharmacology, West Virginia University, Morgantown, WV, USA
* To whom correspondence should be addressed. E-mail: phe{at}hsc.wvu.edu.
The objective of this study was to investigate whether leukocyte adhesion and/or emigration are critical steps in increased microvessel permeability during acute inflammation. To conduct this study, we combined autologous blood perfusion with single microvessel perfusion technique, which allows microvessel permeability to be measured precisely after the endothelium has interacted with blood-borne stimuli. Experiments were carried out in intact venular microvessels in rat mesenteries. Firm attachment of leukocytes to endothelial cells was induced by intravenous injection of TNF-
(3.5 µg/kg) and resuming autoperfusion in a pre-cannulated microvessel. Leukocyte emigration was facilitated by superfusion of formyl-Met-Leu-Phe-OH (fMLP). Microvessel permeability was measured as hydraulic conductivity (Lp) or solute permeability coefficient to TRITC-labeled -lactalbumin before and after leukocyte adhesion and emigration in individually perfused microvessels. We found that perfusion of a microvessel with TNF- did not affect basal microvessel permeability, but intravenous injection of TNF- caused significant leukocyte adhesion. However, the significant leukocyte adhesion and emigration did not cause corresponding increases in either Lp or solute permeability. Thus, our results suggest that leukocyte adhesion and emigration do not necessarily increase microvessel permeability and the mechanisms that regulate the adhesion process act independently from mechanisms that regulate permeability. In addition, silver staining of endothelial boundaries demonstrated that leukocytes preferentially adhered at the junctions of endothelial cells. The appearance of the silver lines indicates that the TNF--induced firm adhesion of leukocyte to microvessel walls did not involve apparent changes in junctional structure of endothelial cells, which is consistent with the results of permeability measurements.
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