Ultrastructural localization and characterization of proteoglycans in human lung alveoli

In order to localize and characterize proteoglycans in human lung alveoli, we have used the cationic dye Cuprolinic Blue according to the critical electrolyte concentration method. After staining, five types of Cuprolinic Blue-positive filaments become apparent: two types in the basement membranes of type I and type II epithelial cells respectively and lying in one or two layers; one type, more scattered, localized in the basement membrane of the endothelial cells and another kind associated with collagen fibrils and separated from each other according to the main banding period (+/- 60 nm) of these fibrils. Finally, there was a type of filament which was only locally present at a variety of places. The basement membrane filaments were sensitive to heparinase, heparitinase, pronase (without prefixation) and nitrous acid treatment, but not to Streptomyces hyaluronidase, neuraminidase, chondroitinase ABC, chondroitinase AC, pronase (after prefixation) and 2.0 M MgCl2 treatment. The basement membrane filaments, therefore, represent heparan sulphate-containing proteoglycans. On the other hand, the collagen fibril associated filaments were sensitive to treatment with heparinase, chondroitinase ABC and pronase (without prefixation), but insensitive to Streptomyces hyaluronidase, neuraminidase, nitrous acid, heparitinase, chondroitinase AC, pronase (after prefixation) and 2.0 M MgCl2 (after prefixation) treatment. These filaments thus represent iduronic acid-rich dermatan sulphate-containing proteoglycans. Several physiological functions for these proteoglycans are discussed.