Brazilin increased [3H]2-deoxyglucose uptake in isolated rat epididymal adipocytes. The fact that calcium may be required for the stimulatory effects of insulin on glucose transport suggests that brazilin might also require calcium for its glucose transport-stimulating action. Changes in the concentration of extracellular calcium had no significant effect on brazilin-induced glucose transport. Nifedipine and verapamil decreased brazilin-induced glucose transport, and quin2-AM abolished the effect of brazilin on glucose transport. A23187, however, showed no effect on brazilin action. 45Ca2+ uptake into adipocytes was not influenced by brazilin treatment, and trifluoperazine significantly inhibited the effect of brazilin on glucose transport. These data suggest that calmodulin and the maintenance of the intracellular calcium concentration, rather than an increase in it, may be essential for the stimulatory action of brazilin on glucose transport.