We used a post-embedding ultrastructural immunogold method to localize osteopontin in human gallbladder epithelial cells. This glycoprotein, originally described in bone but recently found to have a much wider distribution in many epithelia and in some mesenchymal cells, was present in the filamentous glycocalyx, small apical cytoplasmic smooth membrane-bound vesicles, large membrane-bound cytoplasmic granules, and in portions of the Golgi complex in gallbladder columnar epithelial cells. These findings suggest that newly synthesized osteopontin is packaged in Golgi-derived granules that release their contents by classical exocytosis from the cell surface. At least a portion of secreted osteopontin remains on the cell surface, where it becomes integrated into the filamentous glycocalyx coating the luminal surface of gallbladder epithelial cells.