The localization and biosynthesis of perlecan, a basement membrane-type heparan sulfate proteoglycan, were studied in developing tooth germs by using murine molars in neonatal and postnatal stages and primary cultured cells of the enamel organ and dental papilla to demonstrate the role of perlecan in normal odontogenesis. Perlecan was immunolocalized mainly in the intercellular spaces of the enamel organ as well as in the dental papilla/pulp or in the dental follicle. By in situ hybridization, mRNA signals for perlecan core protein were intensely demonstrated in the cytoplasm of stellate reticulum cells and in dental papilla/pulp cells, including odontoblasts and fibroblastic cells in the dental follicle. Furthermore, the in vitro biosyntheses of perlecan core protein by the enamel organ and dental papilla/pulp cells were confirmed by immunofluorescence, immunoprecipitation, and reverse transcriptase-polymerase chain reaction. The results indicate that perlecan is synthesized by the dental epithelial cells and is accumulated in their intercellular spaces to form the characteristic stellate reticulum, whose function is still unknown.