Nitric oxide is a ubiquitous cellular messenger that plays a role in a variety of biological mechanisms. Within the central nervous system, it is formed in glia and neuronal cells and can diffuse away from its site of origin to modulate membrane conductances, neurotransmitter release, behavior, or to control the blood flow within its region of action. Nitric oxide has been shown to modulate gap junction conductance in the retina and cortex, an action it shares with a number of neurotransmitters. In this study, we found that the activation of cortical afferents increased dye coupling between rat neostriatal neurons recorded in vitro. This effect was mimicked by a nitric oxide donor and prevented by a nitric oxide synthase inhibitor, suggesting that activation of corticostriatal fibers may open putative gap junctions in the striatum via release of nitric oxide. This is the first report showing that synaptic modulation of gap junctions can be mediated by nitric oxide, and may provide a mechanism to explain cortical modulation of subcortical pattern formation within this highly integrative structure.