Fluid phase  endocytosis of [125I]iodixanol in rat liver parenchymal, endothelial and Kupffer cells.
Endocytosis of [125I]iodixanol was studied in vivo and in vitro in rat liver cells to determine fluid phase endocytic activity in different liver cells (hepatocytes, Kupffer cells and endothelial cells). The Kupffer cells were more active in the uptake of [l25I]iodixanol than parenchymal cells or endothelial cells. Inhibition of endocytic uptake via clathrin-coated pits (by potassium depletion and hypertonic medium) reduced uptake of [125I]iodixanol much more in Kupffer cells and endothelial cells than in hepatocytes. To gain further information about the importance of clathrin-mediated fluid phase endocytosis, the expression of proteins known to be components of the endocytic machinery was investigated. Using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, endothelial cells and Kupffer cells were found to express approximately fourfold more rab4, rab5 and rab7 than parenchymal cells, while clathrin was expressed at a higher level in endothelial cells than in Kupffer cells and hepatocytes. Using electron microscopy it was shown that liver endothelial cells contained approximately twice as many coated pits per membrane unit than the parenchymal and Kupffer cells, thus confirming the immunoblotting results concerning clathrin expression. Electron microscopy on isolated liver cells following fluid phase uptake of horseradish peroxidase (HRP) showed that HRP-containing organelles had a different morphology in the different cell types: In the liver endothelial cells HRP was in small, tubular endosomes, while in Kupffer cells HRP was mainly found in larger structures, reminiscent of macropinosomes. Parenchymal cells contained HRP in small vacuolar endosomes with a punctuated distribution. In conclusion, we find that the Kupffer cells and the endothelial cells have a higher pinocytic activity than the hepatocytes. The hepatocytes do, however, account for most of the total hepatic uptake. The fluid phase endocytosis in liver endothelial cells depends mainly on clathrin-mediated endocytosis, while the parenchymal cells have additional clathrin-independent mechanisms that may play an important role in the uptake of plasma membrane components. In the Kupffer cells the major uptake of fluid phase markers seems to take place via a macropinocytic mechanism.
Kjeken
R
R
Division of Molecular Cell Biology, Biology Institute, University of Oslo, Norway.
Mousavi
S A
SA
Brech
A
A
Gjøen
T
T
Berg
T
T
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002469:Cell Separation; D002478:Cells, Cultured; D002966:Clathrin / Q000378:metabolism; D003287:Contrast Media / Q000493:pharmacokinetics; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D004727:Endothelium / Q000378:metabolism* / Q000648:ultrastructure; D020558:GTP Phosphohydrolases / Q000096:biosynthesis; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D006735:Horseradish Peroxidase / Q000276:immunology / Q000378:metabolism; D007728:Kupffer Cells / Q000378:metabolism* / Q000648:ultrastructure; D008099:Liver / Q000166:cytology*; D008297:Male; D008854:Microscopy, Electron; D051381:Rats; D017208:Rats, Wistar; D012462:Saline Solution, Hypertonic / Q000378:metabolism; D013997:Time Factors; D014018:Tissue Distribution; D014283:Triiodobenzoic Acids / Q000493:pharmacokinetics*
D000906:Antibodies; D002966:Clathrin; D003287:Contrast Media; D012462:Saline Solution, Hypertonic; D014283:Triiodobenzoic Acids; D006735:Horseradish Peroxidase; D020558:GTP Phosphohydrolases; C044834:iodixanol