Interplay of Light, Melatonin, and Circadian Genes in Skin Pigmentation Regulation.

Circadian regulation of skin pigmentation is essential for thermoregulation, ultraviolet (UV) protection, and synchronization of skin cell renewal. This regulation involves both cell-autonomous photic responses and non-cell-autonomous hormonal control, particularly through melatonin produced in a light-sensitive manner. Photosensitive opsins, cryptochromes, and melatonin regulate circadian rhythms in skin pigment cells. We studied light/dark cycles and melatonin coordination in melanin synthesis and cell proliferation of Xenopus laevis melanophores. In vivo, tadpole pigmentation shows robust circadian regulation mainly hormone-driven, in that isolated melanophores respond strongly to melatonin but only slightly to light. Melanophore proliferation is faster in the dark and slower with melatonin as compared to a 12/12 light/dark cycle. Expression of circadian core genes (clock, bmal1, per1, per2, per3, cry1, cry2, and cry4) in melatonin-treated cells during the light phase mimics dark phase expression. Overexpression of individual Crys did not affect melanization or cell proliferation, likely due to their cooperative actions. Melanin synthesis was inhibited by circadian cycle deregulation through (a) pharmacological inhibition of Cry1 and Cry2 degradation with KL001, (b) continuous light or dark conditions, and (c) melatonin treatment. Our findings suggest that circadian cycle regulation, rather than proliferative capacity, alters melanization of melanophores.