Melanin is usually produced in melanocytes; however, our histologic research indicates that brownish black persistent pigmentation occurs at the basal layer and, partially, at the suprabasal layer of the epidermis, outside of melanocytes, in skin exposed to high doses of ultraviolet (UV) A radiation. This pigmentation remains for several weeks. We examined whether colorless melanogenic precursors participate in this UVA-induced persistent pigmentation. Among them, 5,6-dihydroxyindole-2-carboxylic acid (DHICA) and its O-methyl metabolite, 6-hydroxy-5-methoxyindole-2-carboxylic acid (6H5MICA), can change into dark-colored pigment upon exposure to UVA, but not UVB, radiation, producing irreversible brownish black pigmentation. These hydroxyindole derivatives readily changed into dark-colored pigment upon UVA irradiation in the presence of hydrogen peroxide in vitro. Histochemical and biochemical studies demonstrated that 6H5MICA accumulates in excised skin specimens and supernatant of cultured human melanocytes. These results suggest that dark-colored pigment is generated directly, outside of melanocytes, in response to UVA exposure, by photooxidation of O-methyl metabolites of DHICA, such as 6H5MICA, at least. We propose that the pigmentation derived from this compound at the basal and suprabasal layers of the epidermis is involved in the mechanism of skin persistent pigmentation without reddening, observed after exposure to sunlight.