Excitatory postsynaptic potentials evoked in rat neostriatal spiny projections neurons were followed by a long (100-300 ms) period of membrane hyperpolarization, followed in turn by a late depolarization. Concomitant with these changes in membrane potential were inhibition and subsequent excitation of spontaneous firing and excitatory activity evoked from substantia nigra and cerebral peduncle, but not from cortical stimulating sites. Thalamic-evoked excitatory activity was sometimes sensitive and sometimes insensitive to this inhibition, which has previously been believed to result from intrinsic inhibitory synaptic activity among neostriatal neurons. In intracellular recordings from neostriatal neurons in urethane anesthetized rats this long-lasting inhibitory response (1) exhibited alterations with intracellularly applied steady currents comparable to those of the EPSP, (2) failed to respond to intracellular injection of chloride ions, (3) was associated with either a decrease or no detectable change in the input conductance of the neurons, and (4) was abolished after lesions that interrupted polysynaptic pathways to neostriatum through intracortical and intrathalamic synaptic circuits. These findings indicate that the long lasting inhibitory portion of the responses of neostriatal neurons arises from a phasic inhibition of tonically active corticostriatal and thalamostriatal neurons and a concurrent decrease in the excitability of polysynaptic pathways converging on neostriatal neurons.