Classically, the Edinger-Westphal nucleus is described as containing neurons controlling accommodation and pupillary constriction via projections to the ciliary ganglion. However, in several species including rat, some Edinger-Westphal neurons have ascending or descending CNS projections suggesting that the Edinger-Westphal nucleus might also have non-ocular functions. To further characterize the function of this nucleus we studied the electrophysiological properties of Edinger-Westphal neurons in a slice preparation from juvenile rats. The position of the Edinger-Westphal nucleus was determined using an immunohistochemical procedure directed at the peptide Urocortin, which is expressed in Edinger-Westphal neurons. Passive and active membrane responses were investigated and two different neuron types were identified. One type had a transient firing response to 400 ms depolarizing current pulses and one type had a repetitive firing response. Transient-firing neurons had an outward rectifying response inhibiting firing, possibly due to slowly inactivating I(D)-like potassium channels since low concentrations (200 microM) of the potassium channel blocker 4-aminopyridine elicited repetitive firing. In all neurons, low threshold Ca(2+) spikes were seen and these were blocked by nickel(II) chloride hexahydrate, suggesting that they are mediated via low voltage-activated Ca(2+) channels. Some biocytin-labeled neurons had axons or axonal collaterals projecting laterally or dorsally, suggesting possible non-ocular targets. In conclusion, the rat Edinger-Westphal nucleus contains two separate types of neurons with distinct electrophysiological properties.