The aim of this study was to further characterize the rapid effects of insulin and the tyrosine phosphatase inhibitor vanadate to amplify cell surface insulin binding capacity in isolated rat adipocytes. The effect of 20 min insulin treatment (1000 µU/ml) was 2- to 3-fold (p < 0.01) when cells were treated in medium containing 5.6 mM D-glucose, but it was totally absent in glucose-free medium. Other carbon energy sources, such as fructose and pyruvate, could only partly substitute for D-glucose, with an approximately 1.5-fold insulin effect. Moreover, inhibiting transmembrane glucose transport with cytochalasin B completely blocked the effect of insulin to enhance cell surface binding. The effect of vanadate was only partly glucose-dependent, since a submaximal effect (1.5- to 2-fold, p < 0.05) was seen also in the absence of glucose. The tyrosine kinase inhibitor genistein markedly blunted the effect of vanadate (from 3- to 4-fold to approximately 2-fold, p < 0.05) also indicating the importance of tyrosine phosphorylation-related mechanisms in the upregulation of cell surface insulin binding. Glycosylation of insulin receptors as a mechanism for this effect appears unlikely since neither the effect of insulin nor that of vanadate was altered by the glycosylation inhibitor tunicamycin. The time course for the insulin effect displayed a long duration (at least 6 h), suggesting a maintenance role of insulin keeping its receptors accessible for ligand binding at the cell surface. In conclusion, the effect of insulin and vanadate to upregulate cell-surface insulin receptors is energy-dependent and to some extent specifically glucose-dependent.
Key words
Insulin - Vanadate - Insulin Receptor - Energy-Dependency - Tyrosine Phosphorylation - Glucose - Rat Adipocytes
