Expression and functional activity of P-glycoprotein in cultured hepatocytes from Oncorhynchus mykiss.
P-glycoproteins encoded by multidrug resistance 1 (mdr1) genes are ATP-dependent transporters located in the plasma membrane that mediate the extrusion of hydrophobic compounds from the cell. Using cultured isolated rainbow trout hepatocytes, we characterized an mdr1-like transport mechanism of the teleost liver. Immunoblots with the monoclonal antibody C219, which recognizes a conserved epitope of P-glycoproteins, revealed the presence of immunoreactive protein(s) of 165 kDa in trout liver and cultured hepatocytes. In trout liver sections, the immunohistochemistry with C219 stained bile canalicular structures. Compounds known to interfere with mdr1-dependent transport (verapamil, vinblastine, doxorubicin, cyclosporin A, and vanadate) all increased the accumulation of rhodamine 123 by hepatocytes. Verapamil, vinblastine, and cyclosporin A decreased the efflux of rhodamine 123 from hepatocytes preloaded with rhodamine 123. By contrast, the substrate of the canalicular cation transporter tetraethylammonium and the inhibitor of the multidrug resistance-associated protein MK571 had no effect on rhodamine 123 transport. The results demonstrate the presence of an mdr1-like transport system in the teleost liver and suggest its function in biliary excretion.
Sturm
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Department of Chemical Ecotoxicology, UFZ Centre for Environmental Research, D-04318 Leipzig, Germany. sturm@beaulieu.rennes.inra.fr
Ziemann
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C
Hirsch-Ernst
K I
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Segner
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H
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002478:Cells, Cultured; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015151:Immunoblotting; D007166:Immunosuppressive Agents / Q000494:pharmacology; D017686:Oncorhynchus mykiss; D011422:Propionates / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D020112:Rhodamine 123 / Q000378:metabolism / Q000493:pharmacokinetics
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000970:Antineoplastic Agents; D002121:Calcium Channel Blockers; D007166:Immunosuppressive Agents; D011422:Propionates; D011804:Quinolines; D020112:Rhodamine 123; C059141:verlukast