Rhodamine exclusion activity in primary cultured turbot (Scophthalmus maximus) hepatocytes |
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| Authors: | Tutundjian Renaud Minier Christophe Le Foll Frank Leboulenger François |
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| Institution: | Laboratoire d'Ecotoxicologie, UPRES-EA 3222, Université du Havre, France. renaud.tutundjian@univ-lehavre.fr |
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| Abstract: | Cellular detoxification by direct processes has been investigated in fish by studying the ability of hepatocytes prepared from juvenile aquarium-reared turbot (Scophthalmus maximus) to actively exclude the fluorescent dye rhodamine B (RB). Cell viability was studied by measurements of non-specific esterase activity using fluorescein diacetate. This revealed that turbot hepatocytes can be cultured for a few days with a viability decreasing to 38% after 24 h. The 24-h cultured cells have been used to study the rhodamine B exclusion activity using confocal laser microscopy. Hepatocytes accumulated the dye in a competitive manner with verapamil, thus suggesting that they express a transport system similar to the P-glycoprotein-mediated multixenobiotic resistance process. Incubation of cells with 1 microM RB and 20 microM verapamil led to a 26% increase of cellular fluorescence as compared to the accumulation in absence of competitor. Rhodamine B accumulated in the whole cytoplasm, with more concentrated areas that might correspond to the lysosomal compartment and the cell membrane. |
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