Quantitative cytochemical analysis by microdensitometry of spontaneous or alpha-MSH-stimulated melanogenesis in B16 melanoma cells cultivated in vitro.

A spontaneous cyclic phenomenon characterized by successive waves of either high proliferative rate or intense melanogenesis is described in non-confluent B16 melanoma cells subcultivated during 2 months (or more). Dopaoxidase activity is quantified in individual cells after L-dopa reaction, by an original method of visible light absorption cytophotometry. A 24-hr treatment with alpha-MSH increases dopa-oxidase activity. This increase is also noted during the following 14 hr, in a fresh medium devoid of alpha-MSH, in which cell proliferation resumes after 24 hr. Phenylthiourea, cycloheximide or actinomycin D inhibit dopaoxidase activity, but also cell proliferation in alpha-MSH pre-stimulated cells. The effects of the two latter agents suggest that de novo synthesis of the enzyme takes place following alpha-MSH treatment.