AbstractAmphibian liver pigment cells, also known as Kupffer cells, contain various amounts of melanin, haemosiderin and lipofuscin. We used different histochemical and ultrastructural methods to analyse and compare the level of hepatic pigmentation and the structure of hepatic pigment cells in the livers of three representatives of the family Proteidae; two subspecies of the hypogean Proteus anguinus (depigmented Proteus a. anguinus and pigmented Proteus a. parkelj) and the epygean Necturus maculosus. Our analysis revealed differences at histological and ultrastructural level. While the percentage of the pigmented area and ultrastructural characteristics are similar in both subspecies of P. anguinus, great differences occur in the amount and structure of the pigment cells between P. anguinus and N. maculosus. Pigment cells are more numerous and compose larger pigmented clusters in P. anguinus. They are structurally more heterogeneous and contain a larger amount of haemosiderin when compared to N. maculosus. Our results confirm a high degree of variation in hepatic pigmentation among different amphibian species. Because many factors influence the level of hepatic pigmentation in poikilotherms, differences among species from different habitats and also among individuals of the same species are expected but are not easily explained. We propose two possible explanations for the large amount of iron present in Proteus anguinus: (1) accumulation of pigments due to the very low metabolic rate and extended lifetime; (2) large iron storage capacity as an adaptation to a low and discontinuous food supply in caves.