The competitive taurine uptake inhibitor 2-guanidinoethane sulfonate (GES) is frequently used to deplete cerebral pools of taurine. Previous work has revealed that extracellular glutamate increases during GES administration. In view of this, accumulation of glutamate in the absence or presence of 10 or 100 microM GES was measured in primary astroglial cultures from the rat cerebral cortex. At 100 microM, GES reduced Km as well as Vmax for glutamate uptake. A similar tendency was seen at 10 microM GES, but this was not statistically significant. The data suggest that GES is an uncompetitive inhibitor of reuptake of glutamate in astrocytes, which may underlie the previously noted GES-induced elevation of extracellular glutamate in vivo.