Glypicans are a family of glycosylphosphatidylinositol (GPI)-anchored cell surface heparan sulfate proteoglycans (HSPGs). The glypican family, which currently includes glypican, the developmentally regulated rat intestinal transcript OCI-5, and cerebroglycan, is characterized by a similar core protein size and almost complete conservation of cysteine residues. By RT-PCR using degenerate oligonucleotide primers based on the sequence homologies, we isolated mouse cDNA encoding a novel member of the glypican family as well as mouse homologues of glypican and OCI-5. The novel molecule, named K-glypican, has a predicted molecular mass of 57.5 kD and potential attachment sites for heparan sulfate chains and a GPI anchor in its COOH-terminal region, like other members of the glypican family. Transfection of an epitope-tagged full-length K-glypican cDNA into MDCK cells demonstrated that K-glypican is indeed expressed as a GPI-anchored HSPG. Northern blot analyses with K-glypican, glypican, and OCI-5 probes demonstrated that kidney and developing brain, and that these three molecules show remarkable patterns of cell type- and developmental stage-specific expression. In situ hybridization revealed that the major sites of K-glypican expression in developing embryo are tubular epithelial cells in the kidney and proliferating neuroepithelial cells in the brain. These results indicate that K-glypican is a novel GPI-anchored HSPG involved in embryonic development.