The role of microfilaments and microtubules in taurocholate uptake by isolated rat liver cells.
The role of microfilaments and microtubules on bile salt transport was studied by investigating the influence of a microfilament and a microtubule inhibitor, cytochalasin B and colchicine, respectively, on taurocholate uptake by isolated hepatocytes in vitro. Hepatocytes were prepared by the enzyme perfusion method and [14C]taurocholate uptake velocity was determined by a filtration assay. Taurocholate uptake obeyed Michaelis-Menten kinetics, maximal uptake velocity and apparent half-saturation constants averaging 0.87 +/0 SD 0.05 nmol . s-1 . 10(-6) cells and 10.9 +/- 1.8 muM, respectively. Cytochalasin B (4.2-420 muM) inhibited taurocholate uptake in a competitive fashion; Ki being 33 +/- 7 muM. At concentrations above 100 muM the compound decreased 36Cl membrane potential and intracellular K+ concentration. Other parameters of cell viability were not affected by cytochalasin B. Colchicine (0.1-1.0 mM), by contrast, inhibited taurocholate uptake non-competitively, Ki being 0.47 +/- 0.07 mM. The inhibition brought about by colchicine was considerably smaller than that induced by cytochalasin B. None of the parameters of cell viability tested was affected by colchicine. These results suggest that microfilaments may be involved in the carrier-mediated hepatocellular transport of bile salts. This could, at least in part, account for cytochalasin B-induced cholestasis. The contribution of the microtubular system, if any, is less important quantitatively. The mechanisms whereby these two components of the cytoskeleton partake in bile salt transport remain to be elucidated.
D000818:Animals; D001692:Biological Transport; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D008870:Microtubules / Q000378:metabolism*; D011188:Potassium / Q000378:metabolism; D051381:Rats; D013656:Taurocholic Acid / Q000378:metabolism*
D003571:Cytochalasin B; D013656:Taurocholic Acid; D011188:Potassium; D003078:Colchicine; D004121:Dimethyl Sulfoxide