Glutamatergic and GABAergic responses of the neostriatal spiny neurons to intrastriatal and cortical stimulation were characterized by intracellular recording in brain slice preparations. This study also demonstrated the role of each response in the spike activity of the spiny neuron. Single neostriatal stimulation induced postsynaptic potentials consisting of multiple components. The early part of the postsynaptic potential, which was isolated by the GABAA antagonist bicuculline methiodide and the N-methyl-D-aspartate antagonist 3-(2-carboxypiperzin-4-yl)-propyl-1-phosphonic acid (CPP), was mainly an alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate receptor-mediated response. Perfusion of magnesium-free medium containing bicuculline methiodide and the AMPA/kainate antagonist 3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline (NBQX) disclosed a large, slow N-methyl-D-aspartate receptor-mediated response. The N-methyl-D-aspartate response in magnesium-containing perfusing medium was small in neurons at the resting membrane potential, but became a significant component when the neurons were depolarized to subthreshold membrane potential. The duration of the N-methyl-D-aspartate response was over 300 ms. The nicotinic antagonists dihydro-beta-erythroidine hydrobromide and mecamylamine failed to change responses to single stimulation. Repetitive intrastriatal stimulation induced a large, long-duration depolarization with action potentials in the spiny neurons. This stimulation-induced response resembles that of the depolarization stage observed in anesthetized animals. Bicuculline methiodide increased the response amplitude. In contrast, CPP reduced the amplitude of the response to the below the spike generation threshold. The CPP-sensitive N-methyl-D-aspartate response was large and lasted several hundred milliseconds after the termination of repetitive stimulation. Responses of the neostriatal neurons to cortical stimulation were similar to those induced after intrastriatal stimulation. CPP greatly reduced both the response amplitude and the number of spikes triggered from the response. Bicuculline methiodide, on the other hand, greatly increased the response amplitude and the number of spikes. The AMPA/kainate response alone, which was isolated by application of bicuculline methiodide and CPP, did not induce sustained depolarization in spiny neurons to repetitive cortical stimulation. Application of NBQX diminished GABAA response to cortical stimulation. This observation indicates that, for neostriatal spiny neurons to respond with GABAA response after cortical stimulation, the AMPA/kainate response must be induced in the GABAergic secondary neurons in the neostriatum. This study indicates that the main synaptic driving forces of neostriatal spiny neurons include AMPA/kainate, N-methyl-D-aspartate and GABAA responses. Although AMPA/kainate response is the main synaptic input, the generation of the action potentials in neostriatal neurons is greatly influenced by both GABAA and N-methyl-D-aspartate responses.