1. We obtained whole cell patch-clamp recordings from visually identified sympathetic preganglionic neurons (SPNs) in thin (200-300 microns) transverse spinal cord slices of neonatal rats (1-14 days postnatal). Exogenous application of glutamate (100 microM), N-methyl-D-aspartate (NMDA; 100 microM), kainate (100 microM), quisqualate (1 microM), and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA; 50 microM) induced inward currents at a holding potential of -30 mV. 2. Excitatory postsynaptic currents (EPSCs) were evoked by electrical stimulation either in the dorsal horn or the lateral funiculus. They reversed at 1.2 +/- 4.6 (SD) mV and could in most cases (49 of 51) be separated into two components. 3. In the presence of DL-2-amino-5-phosphonovalerate (10-40 microM) the current-voltage (I-V) relationship of the remaining EPSC was linear. When stimulated in the lateral funiculus, its rise time (10-90%) and the time constant of the monoexponential decay were 1.6 +/- 1.0 and 5.5 +/- 2.7 ms, respectively. By contrast, when stimulated in the dorsal horn, this component had a rise time (10-90%) of 3.0 +/- 0.8 ms and a decay time constant of 13.7 +/- 7.6 ms. 4. We studied the NMDA receptor-mediated component of the EPSCs after superfusion of 6-cyano-7-nitroquinoxaline-2,3-dione (5 microM). The I-V relationship of this component had a region of negative slope conductance between -30 and -80 mV, which was abolished in Mg(2+)-free saline. The rise time (10-90%) ranged from 3.3 to 9.5 ms and the decay was biexponential. Both decay time constants increased with depolarization. Mg(2+)-free saline reduced this voltage sensitivity. 5. At a membrane potential of -80 mV and in 1 mM extracellular Mg2+, the NMDA receptor-mediated component represented 74.8 +/- 11.2% of the total charge carried by the EPSCs evoked by stimulation in the dorsal horn. In contrast, when stimulated from the lateral funiculus, 28.9 +/- 18.9% of the total charge carried during the EPSC was mediated by the NMDA receptor-mediated component. The contribution of the NMDA receptor-mediated component increased in both cases with depolarization. In addition, in 2 of 18 SPNs the EPSC evoked in the dorsal horn was exclusively carried by NMDA receptors. 6. We conclude that L-glutamate or a related substance mediates the fast excitatory input onto SPNs. Viscerosomatic and supraspinal inputs form synapses with different topographical locations on the SPN.(ABSTRACT TRUNCATED AT 400 WORDS)