Wortmannin-sensitive trafficking steps in the endocytic pathway in rat liver endothelial cells.
Liver endothelial cells (LECs) play an important homoeostatic role by removing potentially harmful macromolecules from blood. The extremely efficient endocytosis in LECs makes these cells an interesting model for the study of the involvement of phosphoinositides in the different steps of the endocytic process. In the present investigation we have studied the effect of wortmannin, an inhibitor of phosphatidylinositol kinases, on uptake, recycling and intracellular transport of (125)I-labelled ovalbumin, which is taken up in LECs via mannose-receptor-mediated endocytosis. Wortmannin was found to inhibit both uptake and degradation of ovalbumin. Further studies indicated that the reduced uptake via the mannose receptor was due both to a reduction of the number of surface receptors and a reduction in the rate of receptor-ligand internalization. Transport of ligand from endosomes to lysosomes was prevented, leading to increased recycling of internalized ligand. Wortmannin treatment released the Rab5 effector EEA1 from the endosomes and caused reduced size of early endosomes.
Kjeken
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University of Oslo, Institute of Biology, Division of Molecular Cell Biology, P.O. Box 1050, Blindern, 0316 Oslo, Norway.
Mousavi
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SA
Brech
A
A
Griffiths
G
G
Berg
T
T
D000730:Androstadienes / Q000494:pharmacology*; D000818:Animals; D001324:Autoantigens / Q000378:metabolism; D003034:Coated Pits, Cell-Membrane / Q000187:drug effects / Q000502:physiology; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D011992:Endosomes / Q000187:drug effects / Q000502:physiology*; D004727:Endothelium / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D007457:Iodine Radioisotopes; D007700:Kinetics; D037181:Lectins, C-Type; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008297:Male; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins / Q000378:metabolism; D010047:Ovalbumin / Q000493:pharmacokinetics; D000081082:Phosphoinositide-3 Kinase Inhibitors; D021381:Protein Transport; D051381:Rats; D017208:Rats, Wistar; D011956:Receptors, Cell Surface / Q000502:physiology*; D033921:Vesicular Transport Proteins; D000077191:Wortmannin
D000730:Androstadienes; D001324:Autoantigens; D007457:Iodine Radioisotopes; D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011956:Receptors, Cell Surface; D033921:Vesicular Transport Proteins; C093846:early endosome antigen 1; D010047:Ovalbumin; D000077191:Wortmannin