Stereological Studies of the Effects of α‐MSH and cAMP on Melanosomes in Melanoma Cells

The effects of alpha-MSH and cAMP on melanosomes in Cloudman S91 melanoma cells were investigated by modern stereological techniques. Cells were cultured for 4 days in medium containing alpha-MSH or cAMP, harvested at 24 hour intervals; some were frozen for melanin assay and the reminder embedded in Epon for light and electron microscopy. Cellular and melanosomal parameters were estimated by new stereological probes. We found that both stimulators induced increases in nuclear volume, cell volume, and the volume fractions and volumes of premelanosomes (V(Vpm,cell), Vpm) and mature melanosomes (V(Vmm,cell), Vmm), and the number of mature melanosomes (Nmm). Both stimulators also caused declines in the volume of individual mature melanosomes (V(Vimm)), the melanin content per mature melanosome unit volume and the melanin content per individual mature melanosome. The increases in the volume of individual premelanosomes and the number of premelanosomes were only induced by cAMP. The effect cAMP on some parameters occurred 24 hours prior to alpha-MSH and was more marked. The response of premelanosomes to the stimulators was more sensitive than mature melanosomes. These results suggest that both stimulators enhanced melanogenesis by increasing the V(Vpm,cell), V(Vmm,cell), Vpm, Vmm and Nmm. The melanogenic level did not depend on the V(Vimm) and melanin concentration in melanosomes. The maturation of premelanosomes was involved in melanogenesis induced by both stimulators, but, de novo synthesis and enlargement of premelanosomes were only stimulated by cAMP. It imply that exogenous cAMP may affect melanosomes, and hence melanogenesis in quantitatively or qualitatively different ways to alpha-MSH.