An immunohistochemical study of endothelial cell heterogeneity in the rat: observations in "en face" Häutchen preparations.

"En face" sheets of endothelium, in which cellular spatial relationships were maintained, were prepared from proximal pulmonary and femoral arteries and aortae of the Wistar rat, in order to visualise patterns of heterogeneity in populations of endothelial cells. These preparations, termed Häutchen, were immunolabelled with antibodies to angiotensin II, endothelin and Factor-VIII-related antigen, and visualised by an avidin/biotin peroxidase complex. Clusters of cells, which accounted for approximately 30% of the total endothelial cell population, and which were positively immunostained for angiotensin II, were found perpendicular to the longitudinal axis of the aorta and femoral artery. Cells in the pulmonary artery were immunonegative for angiotensin II. The majority of cells in all three vessels were immunopositive for endothelin; groups of intensely stained cells were present in both the femoral artery and aorta, but not in the pulmonary artery. Immunoreactivity to Factor-VIII-related antigen was heterogeneous, with intensely stained amorphous patches of endothelial cells present in the femoral artery and aorta. Häutchen preparations present an opportunity for the investigation of endothelial cell heterogeneity, both within and between vessels; this may provide a basis for the interpretation of the heterogeneity of endothelium-dependent responses in vessels of differing origin.