Endocytosis and Na+/solute cotransport in renal epithelial cells.
Endocytic uptake of [3H]sucrose and lucifer yellow, markers for fluid-phase endocytosis, was studied in cultures of the renal epithelial cell lines LLC-PK1 and OK. Endocytosis in LLC-PK1 cells was inhibited when the cells were grown in the presence of gentamicin (1 mg/ml) for 4 days or when the cells were treated with concanavalin A (1 mg/ml) for 5 h. These changes occurred without perturbation of intracellular Na+ and K+ content, indicating that the cells maintained normal ion gradients. The inhibition of endocytosis was accompanied by marked increases in the apparent Vmax for Na+-dependent cell uptake of solutes such as Pi and L-alanine. The apparent Km was unchanged. In contrast, treatment of OK cells with concanavalin A produced marked stimulation of endocytosis and inhibition of the Na+-dependent uptake of Pi and L-glutamate. These changes occurred in the absence of changes in intracellular Na+ and K+ content. Neither gentamicin nor concanavalin A had a direct effect on Na+/solute cotransport in these cell lines. The changes in Na+/Pi cotransport induced by concanavalin A in both LLC-PK1 and OK cells were blocked by keeping the cells at 4 degrees C during exposure to the lectin, suggesting that endocytosis may be part of the mechanism which mediates the changes in solute uptake. The reciprocal relationship between the changes in endocytosis and the changes in Na+/solute cotransport is consistent with the possibility that the number of Na+/solute cotransporters present in the plasma membrane may be altered by an increase or decrease in the rate of membrane internalization by endocytosis. The Vmax changes in Na+/solute cotransport provide indirect support for this conclusion.
Kempson
S A
SA
Department of Physiology, Indiana University School of Medicine, Indianapolis 46202-5120.
Ying
A L
AL
McAteer
J A
JA
Murer
H
H
D000409:Alanine / Q000378:metabolism; D001692:Biological Transport; D002460:Cell Line; D003208:Concanavalin A / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D004848:Epithelium / Q000378:metabolism; D005839:Gentamicins / Q000494:pharmacology; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D007546:Isoquinolines / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D010710:Phosphates / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism* / Q000494:pharmacology; D013395:Sucrose / Q000378:metabolism
D005839:Gentamicins; D005971:Glutamates; D007546:Isoquinolines; D010710:Phosphates; D003208:Concanavalin A; D018698:Glutamic Acid; D013395:Sucrose; C017475:lucifer yellow; D012964:Sodium; D000409:Alanine; D011188:Potassium