The lumbosacral spinal network controling penile erection is activated by information from peripheral and supraspinal origins. We tested the hypothesis that glutamate, released by sensory afferents from the genitals, activates this pro-erectile network. In anesthetized intact and T8 spinalized (i.e. freed from supra-spinal inhibition) male rats, the parameters of electrical stimulation of the dorsal penile nerve (DPN) that elicited intracavernous pressure (ICP) rises were determined. In T8 spinalized rats, DPN stimulations were applied in the presence of D(-)-2-amino-5-phosphonopentanoic acid (D-AP5), a competitive NMDA receptor antagonist, or of 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulphonamide (NBQX), a AMPA/kainate receptor antagonist, injected intrathecally at the lumbosacral level. Both antagonists, alone or in combination, dose-dependently decreased the ICP rise and increased its latency. In conscious rats, reflexive erections were depressed by D-AP5 and NBQX, as revealed by an increased latency of the first erection and by decreases of the number of rats displaying erections, of the number of erection clusters and of the number of erections per cluster. In anesthetized rats, the combined administration of the glutamatergic agonists N-methyl-D-aspartic acid (NMDA) and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) elicited ICP rises in the absence of DPN stimulation. In contrast, both agonists moderately decreased the ICP rise elicited by DPN stimulation but did not affect its latency. These results support our hypothesis that glutamate, released upon stimulation of the genitals and acting at AMPA and NMDA receptors, is a potent activator of the spinal proerectile network.