Direction selectivity is a receptive field property displayed by neurons throughout the visual system. Previous experiments have concentrated on the role of lateral connections that use gamma-aminobutyric acid and acetylcholine. We have examined the role of excitatory amino acid receptors on direction-selective ganglion cell function in the rabbit retina. Application of the quinoxalines, a group of kainate/alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor antagonists, selectively blocked the directional-selectivity mechanism, leaving cells responsive to both directions of movement. In contrast, direction selectivity was unaffected by N-methyl-D-aspartate receptor antagonists or L-2-amino-4-phosphonobutyric acid. Large reductions in acetylcholine release by starburst amacrine cells appear to parallel losses of direction selectivity observed in the quinoxalines. These results shed additional insights into the mechanism of direction selectivity.