We performed a biochemical and histochemical study of sulfated glycosaminoglycans in the tissues of the ascidian Styela plicata. A highly sulfated dermatan sulfate and a heparin-like polymer, identified by incubation with specific lyases, occur at different concentrations in intestine, heart, pharynx, and cloak. Dermatan sulfate prevails in the pharynx, whereas the heparin-like polymer abounds in the intestine. Staining of tissues sections with the cationic dye 1,9-dimethylmethylene blue before and after incubation with specific lyases revealed that the dermatan sulfate occurs in the extracellular matrix, while the heparin-like polymer is located within cytoplasmic granules of cells in the lumen of intestine and pharynx. The dermatan sulfate has a similar disaccharide composition in all tissues studied, whereas the heparin-like polymer differs in sulfate content. A direct relationship between sulfate content of the heparin-like polymer and antithrombin activity was observed. Analysis of the repeating disaccharide units of the heparin-like polymer indicates the presence of relatively high amounts of the disulfated disaccharide namely DeltaUA-1-->4-GlcN(SO(4))-(6SO(4)), which may suggest the occurrence in ascidians of regulatory biosynthetic mechanisms different from those observed for heparin in mammals.