An immunofluorescence study of the expression of keratin 14 (K14) during the formation of filiform papillae was performed and the progress of keratinization of the epithelium of the rat tongue was monitored on semi-ultrathin sections by laser-scanning microscopy. Differential interference contrast (DIC) images were also examined to provide details of histology and cell morphology. No cells with immunoreactivity specific for K14 were detected on the lingual epithelium of foetuses on embryonic days 12 and 16 (E12 and E16), when the lingual epithelium was composed of a single layer or several layers of cuboidal cells. Immunoreactivity specific for K14 was detected first on basal and suprabasal keratinocytes of the dorsal epithelium of the tongue of new-borns on postnatal day 0 (P0) and was conspicuous in juveniles on P14. The immunoreactivity was particularly strong on the basal and suprabasal keratinocytes along the connective tissue papillae. The immunoreactivity extended over the entire cytoplasm but was not detected in the nucleus. The lingual epithelium was composed of stratified squamous cells and the rounded rudiments of filiform papillae were compactly arranged at equal intervals, for the most part, and the spaces between them were narrow and indistinct. Immunostaining of K14 was distinct on basal and suprabasal keratinocytes of the filiform papillar area of tongues of juveniles on P21, when the filiform papillae were conical. The spaces between them were relatively wide and, as a result, interpapillar cell columns were clearly visible. Immunoreactivity specific for K14 in the basal and suprabasal keratinocytes of the interpapillar cell columns was recognizable but was weaker than that in cells of papillar cell columns. The thickness of the epithelium in papillar and interpapillar areas increased gradually with the development of filiform papillae. However, sizes of basal and suprabasal keratinocytes remained almost unchanged during this process. These results suggest that the basal and suprabasal keratinocytes of the filiform papillar area proliferate with the initiation of the morphogenesis of filiform papillae and the keratinization of the epithelium. In addition, it appears that, after P14, the basal and suprabasal keratinocytes of the interpapillar area proliferate to supply the keratinocytes of the expanding interpapillar regions. © 2003 Elsevier Science Ltd. All rights reserved.