title: Glucose release from GLUT2-null hepatocytes: characterization of a major and a minor pathway.
abstract: We previously reported that glucose can be released from GLUT2-null hepatocytes through a membrane traffic-based pathway issued from the endoplasmic reticulum. Here, we further characterized this glucose release mechanism using biosynthetic labeling protocols. In continuous pulse-labeling experiments, we determined that glucose secretion proceeded linearly and with the same kinetics in control and GLUT2-null hepatocytes. In GLUT2-deficient hepatocytes, however, a fraction of newly synthesized glucose accumulated intracellularly. The linear accumulation of glucose in the medium was inhibited in mutant, but not in control, hepatocytes by progesterone and low temperature, as previously reported, but, importantly, also by microtubule disruption. The intracellular pool of glucose was shown to be present in the cytosol, and, in pulse-chase experiments, it was shown to be released at a relatively slow rate. Release was not inhibited by S-4048 (an inhibitor of glucose-6-phosphate translocase), cytochalasin B, or progesterone. It was inhibited by phloretin, carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone, and low temperature. We conclude that the major release pathway segregates glucose away from the cytosol by use of a membrane traffic-based, microtubule-dependent mechanism and that the release of the cytosolic pool of newly synthesized glucose, through an as yet unidentified plasma membrane transport system, cannot account for the bulk of glucose release.
authors.affiliation: Institute of Pharmacology and Toxicology, University of Lausanne, CH-1005 Lausanne, Switzerland.
authors.forename: Masaya
authors.initials: M
authors.lastname: Hosokawa
authors.forename: Bernard
authors.initials: B
authors.lastname: Thorens
mesh_terms: D000818:Animals; D017920:Antiporters; D002118:Calcium / Q000378:metabolism; D002250:Carbon Radioisotopes; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D003470:Culture Media; D003571:Cytochalasin B / Q000494:pharmacology; D047630:Depsipeptides; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004734:Energy Metabolism; D005947:Glucose / Q000378:metabolism*; D051273:Glucose Transporter Type 2; D019298:Glucose-6-Phosphate / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008387:Marine Toxins; D051379:Mice; D018345:Mice, Knockout; D009004:Monosaccharide Transport Proteins / Q000172:deficiency* / Q000235:genetics / Q000502:physiology; D015739:Nocodazole / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000494:pharmacology; D010693:Phloretin / Q000494:pharmacology; D010770:Phosphotransferases / Q000037:antagonists & inhibitors; D011374:Progesterone / Q000494:pharmacology; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: D017920:Antiporters; D002250:Carbon Radioisotopes; D003470:Culture Media; D047630:Depsipeptides; D051273:Glucose Transporter Type 2; D008387:Marine Toxins; D009004:Monosaccharide Transport Proteins; D010080:Oxazoles; D010456:Peptides, Cyclic; C503532:Slc37a4 protein, mouse; D014475:Uncoupling Agents; C111261:glucose 6-phosphate(transporter); C057531:jasplakinolide; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D003571:Cytochalasin B; D011374:Progesterone; D019298:Glucose-6-Phosphate; C059041:calyculin A; D002784:Cholesterol; D010770:Phosphotransferases; D005947:Glucose; D010693:Phloretin; D015739:Nocodazole; D002118:Calcium