Visualization and quantification of transmural concentration profiles of macromolecules across the arterial wall

Transport parameters that describe a macromolecule entering the arterial wall from plasma can be obtained from concentration profiles of the labeled macromolecule entering the tissue. A new technique has been developed for measuring such concentration profiles, which offers spatial resolution superior to methods that measure profiles of radiolabeled macromolecules by serially sectioning tissue in planes parallel to the endothelium. In addition, this new method preserves cellular organization and tissue structure and permits measurement of concentration profiles underlying focal endothelial injuries or vascular lesions. The technique quantifies the concentration of a protein by measuring associated peroxidase activity. Although the present study was performed using horseradish peroxidase (HRP), the same principles can be applied to other macromolecules linked to HRP or microperoxidase. The colored reaction product of HRP was detected in transverse aortic sections using an image processing system. In the present study, profiles obtained by this new method were validated by comparison with HRP concentration profiles in rat aortas obtained by a serial slicing technique using radiolabeled HRP. We used the technique to measure high-resolution HRP concentration profiles in the intima and media of normal animals. These concentration profiles suggest that the internal elastic lamina acts as a major barrier to transport of macromolecules across the wall of the normal rat aorta. The new method should allow concentration profiles for macromolecules to be quantified in tissue surrounding vessels in the microcirculation, within the thickened intima of large vessels, and across coronary artery walls.