Immunostaining of adult human skin shows that the small dermatan sulphate proteoglycan decorin is abundant in the whole dermal layer but absent from the epidermis. In the papillary layer adjacent to the dermal-epidermal border, more decorin was detected than in the reticular layer of the dermis. Expression of decorin mRNA by cells in the papillary dermis could also be shown by in situ hybridization. In contrast, biglycan, another small chondroitin sulphate/dermatan sulphate proteoglycan, is found only at the dermal-epidermal border. Therefore the biosynthesis of these two proteoglycans by papillary and reticular fibroblasts from two different donors was compared in tissue culture. Papillary fibroblasts secrete up to 5.9 times more decorin than reticular fibroblasts, while the amounts of cell-associated decorin in both cell types are similar. By Northern blot analysis as well as by in situ hybridization it was shown that papillary fibroblasts contain more mRNA coding for decorin than do reticular cells. In addition, no mosaic pattern of decorin expression was found in the cultured cells. The expression and synthesis of biglycan compared with decorin was about 10 times lower and did not show any significant differences for the two cells types. The kinetics of secretion and the rate of endocytosis of decorin were similar for both types of fibroblasts. These results were found with fibroblasts between the 9th and 15th passage from a newborn subject as well as from a 78-year-old donor, indicating that the pattern of decorin synthesis is not age-dependent in the range investigated. These results further show that fibroblasts from different layers of the dermis have a specific pattern of synthesis of small chondroitin sulphate/dermatan sulphate proteoglycans, and they also maintain these patterns in cell culture.