Melanogenic response of the Kupffer cells of Rana esculenta L to melanocyte stimulating hormone

We have previously shown that Kupffer cells (KCs) of Rana esculenta L. possess melanogenic ability. The melanogenic enzyme activities in these cells are different from those described in skin melanocytes, and very little is known about their regulation by extracellular signalling molecules. In order to study this regulation, we analysed the effects of NDP-MSH on the levels of expression of the tyrosinase gene and on dopa-oxidase activity, using primary cultures of KCs. Incubation of the cells with NDP-MSH increases tyrosinase gene transcription, within the first 24 h of stimulation. To gain insight into the signalling mechanism involved in the cell response to the hormone, KCs in culture were incubated with IBMX or forskolin. These agents mimic the effects of alpha-MSH on melanocytes by increasing the intracellular level of cAMP. The experimental results showed that while the hormonal treatment always activated the KC tyrosinase system, treatment with IBMX or forskolin never did. Therefore, in KCs the tyrosinase-stimulating action of NDP-MSH was not mimicked by cAMP elevating agents. Assays of cAMP levels in cells stimulated with NDP-MSH demonstrated that the hormone does not produce significant increases in intracellular cAMP. On the contrary, forskolin produced significant increases in cAMP starting from 30 min of incubation. These results suggest that tyrosinase induction by melanocortins in KCs is not mediated by the cAMP pathway, and highlight the existence of substantial differences in the hormone signal transduction mechanisms between amphibian KCs and melanocytes or melanoma cells.