Three unrelated perturbations similarly uncouple fluid, bulk-membrane, and receptor endosomal flow in rat fetal fibroblasts.
We have compared the effects of three perturbations (treatment with 2 microM monensin, potassium depletion, and incubation in 0.35 M NaCl) on recycling of internalized fluid-phase, bulk-membrane, and receptor-mediated tracers in rat fetal fibroblasts. Monensin accelerated 2-fold the regurgitation of the fluid-phase tracer horseradish peroxidase (HRP), as previously described in these cells after potassium depletion or upon incubation in hypertonic medium (1), and all treatments severely inhibited transfer of HRP from endosomes to lysosomes. In comparison, recycling of (6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]hexanoyl glucosylsphingosine (C6-NBD-GlcCer), a fluorescent lipid used as a bulk-membrane probe, was not significantly affected while transferrin recycling was slowed down 2-fold. The striking similarities of these unrelated perturbations in their distinct effects on fluid, bulk-membrane, and receptor addressing point to common targets regulating these mechanisms.
Cupers
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P
Cell Biology Unit, University of Louvain Medical School, Brussels, Belgium.
Veithen
A
A
Hoekstra
D
D
Baudhuin
P
P
Courtoy
P J
PJ
D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D004622:Embryo, Mammalian; D004705:Endocytosis; D011992:Endosomes / Q000502:physiology*; D005347:Fibroblasts / Q000187:drug effects* / Q000502:physiology* / Q000648:ultrastructure; D006207:Half-Life; D006735:Horseradish Peroxidase / Q000378:metabolism; D007424:Intracellular Fluid / Q000502:physiology; D008985:Monensin / Q000494:pharmacology*; D011188:Potassium / Q000008:administration & dosage*; D051381:Rats; D011990:Receptors, Transferrin / Q000378:metabolism; D012462:Saline Solution, Hypertonic; D012965:Sodium Chloride / Q000008:administration & dosage / Q000494:pharmacology*; D014168:Transferrin / Q000378:metabolism
D011990:Receptors, Transferrin; D012462:Saline Solution, Hypertonic; D014168:Transferrin; D012965:Sodium Chloride; D008985:Monensin; D006735:Horseradish Peroxidase; D011188:Potassium