A large body of evidence suggests that retinoids modulate the phenotypic expression of epithelial cells of skin and mucous membranes. The purpose of this study was to investigate the effect of retinoic acid on keratinocyte-mediated collagen breakdown. Keratinocytes derived from the ventral surfaces of the tongues of 4-6 week-old male Wistar rats were established in culture under conditions which are restrictive to growth of fibroblasts, and they were eventually cloned by limiting dilution. The cells were seeded (100,000 cells/cm2) in dishes coated with 3H-labeled, reconstituted type I collagen fibrils and incubated in serum-free medium over a 3-5 day period. Dissolution of the collagen fibrils was monitored by the release of radioactivity to the culture medium. Unstimulated cells metabolized the collagen rather slowly, but addition of retinoic acid in concentrations from 10(-6)M to 10(-8)M resulted in marked acceleration of the degradative process, with complete solubilization of the collagen fibrils in four or five days. The effect of 10(-6)M retinoic acid was of the same order of magnitude as that obtained by addition of a proteolytic activating system either in the form of plasmin or of plasminogen, which is converted to catalytic plasmin by endogenous activators. The effects of retinoic acid and plasminogen/plasmin, however, were not additive. Keratinocytes rendered vitamin A-deficient by cultivation in sera from deficient rats were clearly less effective in degrading the collagen substrate than were "sufficient" cells. Addition of retinoic acid (10(-7)M) enhanced collagen breakdown in both sets of cultures and partially restored the collagenolytic activity of the deficient cells.