Iodoacetate action on endocytic uptake of different fluid-phase markers by OK renal epithelial cells.
When grown in monolayer culture, OK cells display endocytic uptake of soluble fluid-phase markers such as lucifer yellow (LY) and horseradish peroxidase (HRP). The response of this process to metabolic inhibitors was characterized in the present study. Inhibition of cell metabolism by cyanide produced a decrease in cell ATP content which was accompanied by a decrease in uptake of both LY and HRP, confirming the energy-dependence of fluid-phase endocytosis in OK cells. Use of iodoacetate also decreased cell ATP content but its action on endocytosis was unexpected. Cell uptake of HRP was decreased by iodoacetate, similar to the effect of cyanide, but there was a marked increase in LY uptake. Additional studies showed that cyanide did not change intracellular Na+ or intracellular K+ and did not interfere with the Na(+)-dependency of Pi uptake. In contrast, iodoacetate produced a marked increase in Na+, a decrease in K+, and abolished the Na(+)-dependency of Pi transport. The latter was due primarily to a 10-fold increase in Na(+)-independent uptake of Pi. These findings suggest, indirectly, that plasma membrane permeability to Na+, K+, Pi, and small molecules such as LY, may be increased by iodoacetate, possibly through its action as an alkylating agent. This mechanism may allow increased cell uptake of LY through a non-endocytic pathway, and may mask the inhibitory action of iodoacetate on endocytic uptake of LY. These additional effects complicate the use of iodoacetate to interrupt endocytosis.
Kempson
S A
SA
Department of Physiology and Biophysics, Indiana University School of Medicine, Indianapolis.
Kunkler
K J
KJ
Murer
H
H
D000818:Animals; D002460:Cell Line; D004705:Endocytosis / Q000187:drug effects*; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D005456:Fluorescent Dyes; D006735:Horseradish Peroxidase / Q000378:metabolism; D007461:Iodoacetates / Q000494:pharmacology*; D019807:Iodoacetic Acid; D007546:Isoquinolines; D007668:Kidney / Q000502:physiology*; D007700:Kinetics; D009893:Opossums; D010710:Phosphates / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D011190:Potassium Cyanide / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism
D005456:Fluorescent Dyes; D007461:Iodoacetates; D007546:Isoquinolines; D010710:Phosphates; C017475:lucifer yellow; D012964:Sodium; D006735:Horseradish Peroxidase; D011190:Potassium Cyanide; D011188:Potassium; D019807:Iodoacetic Acid