Accumulation of some hexoses in adipocytes: properties of the system

Adipocytes accumulate 2-deoxy-D-glucose (2-DG) against significant chemical gradients (J. Foley and J. Gliemann. Biochim. Biophys. Acta 648: 100-106, 1981). The specificity of this accumulation process was examined. Isolated rat adipocytes incubated at 37 degrees C with 0.1 mM hexoses accumulated free 2-deoxy-D-galactose (2-DGal) or D-galactose (Gal) against steady-state gradients of 5.5 +/- 1.2 and 2.5 +/- 0.7, respectively. Like 2-DG, both 2-DGal and Gal are substrates for phosphorylation. Insulin and insulin-mimetic agents increased steady-state accumulations of Gal, 2-DGal, and free and phosphorylated 2-DG (100 nM insulin increased free 2-DG from 2.1 +/- 0.2 to 6.6 +/- 0.3 mM; external 2-DG = 0.1 mM). Removal of extracellular calcium or sodium or the presence of A23187 or ouabain failed to inhibit 2-DG accumulation. Plasma membrane permeabilization induced by either digitonin or high-voltage discharge produced a loss of cellular 2-DG and 2-DG-phosphate without affecting 3-O-methyl-D-glucose equilibration. The data indicate that neither transmembrane ionic gradients nor intracellular compartmentation suffice as explanations of the mechanism of the accumulation process. The possible role of phosphorylation in the process of hexose accumulation is discussed.