Rhodamine exclusion activity in primary cultured turbot (Scophthalmus maximus) hepatocytes.
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.
Tutundjian
Renaud
R
Laboratoire d'Ecotoxicologie, UPRES-EA 3222, Université du Havre, France. renaud.tutundjian@univ-lehavre.fr
Minier
Christophe
C
Le Foll
Frank
F
Leboulenger
François
F
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000494:pharmacology*; D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival; D018432:Drug Resistance, Multiple; D005412:Flatfishes / Q000502:physiology*; D005456:Fluorescent Dyes / Q000493:pharmacokinetics*; D022781:Hepatocytes / Q000473:pathology*; D012235:Rhodamines / Q000493:pharmacokinetics*; D015262:Xenobiotics / Q000378:metabolism / Q000494:pharmacology*
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D005456:Fluorescent Dyes; D012235:Rhodamines; D015262:Xenobiotics; C029773:rhodamine B