Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose or L-fucose concentrations. Chronic exposure of neuroblastoma cells to 30 mM glucose or 30 mM L-fucose caused a decrease in ouabain-sensitive and veratridine-stimulated 22Na+ uptake compared with cells cultured in unsupplemented medium. The Na+ current, determined by using whole-cell configuration of the patch clamp, was also decreased in these cells. Tetrodotoxin (3 microM), which blocked whole cell Na+ currents, also blocked veratridine-stimulated 22Na+ accumulation. Culturing cells in medium containing 30 mM fructose as an osmotic control had no effect on Na+ flux. Specific [3H]saxitoxin binding was not affected by 30 mM glucose or 30 mM L-fucose compared with cells grown in unsupplemented medium, suggesting that the number of Na+ channels was not decreased. These studies suggest that exposing cultured neuronal cells to conditions that occur in the diabetic milieu alters Na+ transport and Na(+)-channel activity.