Melanogenesis and evidence for melanosome transport to the plasma membrane in a CD83 teleost leukocyte cell line

Abstract

Visceral organs of ectothermic vertebrates harbour melanin-containing leukocytes termed melanomacrophages. These cells are thought to participate in immune reactions and free-radical trapping. In teleosts, the melanin-producing ability of melanomacrophages has hitherto not been confirmed by molecular techniques. Here, a leukocyte marker and the apparatus for melanosome production and transport were investigated in an Atlantic salmon (Salmo salar) pronephros-derived mononuclear leukocyte (SHK-1) cell line. The SHK-1 cells expressed transcripts specific for a mammalian CD83 homologue, a standard surface marker for activated or differentiated dendritic cells, and dopachrome tautomerase/tyrosinase-related protein-2, a melanocyte specific enzyme essential for melanin production. Reduction potential of melanin or its precursors was demonstrated histochemically after prolonged cultivation. Ultrastructural investigations revealed tyrosinase and acid phosphate activity in identical organelles and BSA-gold co-localized with multilamellar melanosomes after 2 h internalization. Apparently, melanosomes were transported and released through periodically occurring tubules fusing with the plasma membrane. Video monitoring revealed filopodia and macropinocytosis. These results showed that the SHK-1 cell line is capable of melanogenesis and melanosome secretion. Melanin-producing cells in teleost pronephros may represent a distinct CD83(+) leukocyte population consisting of phylogenetically relict multifunctional cells. This is the first report of a melanin-producing leukocyte cell-line.