Effect of glucocorticoids on hexose transport in rat adipocytes. Evidence for decreased transporters in the plasma membrane.
Glucocorticoids are known to rapidly inhibit glucose transport when added to isolated rat adipocytes. To determine whether this inhibition of transport persists following isolation of the plasma membranes, adipocytes were incubated in the absence or presence of a maximally inhibitory concentration of dexamethasone, a synthetic glucocorticoid, and plasma membrane vesicles were prepared. D-Glucose uptake into vesicles from steroid-treated cells was inhibited by an average of 40%. The ability of dexamethasone to inhibit transport depended upon pretreatment of cells with hormone prior to membrane isolation. Furthermore, the decreased rate of transport was prevented by the simultaneous addition to the cell of actinomycin D or cycloheximide with dexamethasone, indicating a requirement for RNA and protein synthesis. The effect of dexamethasone on glucose transport was further investigated using our recently developed cytochalasin B affinity-labeling protocol to identify the transporter on sodium dodecyl sulfate-polyacrylamide gels. A peak of radioactivity having Mr = 54,000 was identified which exhibited the properties expected for the glucose transporter, in that label incorporation was prevented by D-glucose and unlabeled cytochalasin B, but not by D-sorbitol or unlabeled cytochalasins A, D, or E. Dexamethasone was found to cause a significant (average 33%) decrease in the amount of labeled transporter in the plasma membrane which was prevented by the simultaneous addition of actinomycin D with dexamethasone to the cells. A similar percentage decrease was not found in a microsomal membrane fraction nor in a total cellular membrane fraction. These results suggest that glucocorticoids may decrease glucose transport in rat adipocytes by selectively decreasing the number of transporters in the plasma membrane.
D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology*; D005947:Glucose / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D009004:Monosaccharide Transport Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D013012:Sorbitol / Q000494:pharmacology
D002352:Carrier Proteins; D009004:Monosaccharide Transport Proteins; D003609:Dactinomycin; D003571:Cytochalasin B; D013012:Sorbitol; D003907:Dexamethasone; D003513:Cycloheximide; D005947:Glucose