The synthesis of glycosaminoglycans (GAG) in a freshwater mussel was studied in organ culture using labeled precursors. The major GAGs synthesized were determined and characterized by chemical and enzymatic methods. They were shown to be heparin and an unusual type of heparan sulfate. Gills produced about 50% of each polymer; mantles synthesized little heparin and mostly the heparan-sulfate-like compound, which is similar to a GAG isolated previously from lobsters. No significant amounts of chondroitin sulfates were present. Histological data showed that the sulfate-labeled GAGs were present mainly in exterior pericellular and basement membrane locations of gills and mantle. That is, they would be in contact with the external aqueous environment, suggesting a potential role in calcium transport and storage.