Occludin and claudins are tetraspan-transmembrane proteins in tight junctions. Maculae occludentes, which are less-developed tight junctions, occur in the granular cell layer of the epidermis. The periderm, which overlies the developing epidermis and functions as a protective layer for the embryo, carries developed tight junctions as observed in simple epithelia. In both periderm and epidermis, occludin is expressed at the cell-cell border. To determine the difference between tight junctions of periderm and epidermis, claudin-6 expression was examined in periderm and epidermis. Immunofluorescence staining showed claudin-6 expression at the cell-cell border of the periderm, but not in the epidermis. Reverse transcription-polymerase chain reaction confirmed that claudin-6 was not expressed in mouse adult skin, whereas immunoelectron microscopy revealed that claudin-6 was localized at tight junctions of the periderm. Furthermore, L fibroblasts with stable expression of exogenous claudin-6 formed developed tight junctions at cell-cell borders. These findings indicate that molecular architecture of tight junctions of the periderm is different from that of the maculae occludentes of the epidermis, and that claudin-6 is important in the formation of tight junctions of the periderm.