Heart rate is normally dominated by the activity of the cardioinhibitory parasympathetic nervous system, while abnormally low levels of parasympathetic cardiac activity have been implicated in many cardiovascular diseases including hypertension, heart failure and sudden cardiac death. In this study we have examined the voltage-gated currents in parasympathetic cardiac neurons that were identified with a retrograde fluorescent tracer in visualized sections (250 microns) of nucleus ambiguus. Depolarization of parasympathetic cardiac neurons to potentials more positive than -50 mV evoked a rapidly activating and inactivating inward current which could be blocked by tetrodotoxin (TTX), although in some neurons up to 10 microM was required for complete block. The voltage-dependent inactivation properties of this Na current showed relatively broad inactivation characteristics, a characteristic of TTX-resistant Na channels. Depolarization also elicited biphasic outward currents, which were separated into a transient IA type K current using the specific channel antagonist 4-aminopyridine and a long-lasting delayed rectified K current. These voltage-gated Na and K currents define the action potential firing patterns of parasympathetic cardiac neurons, such as frequency adaptation and spike delay, and also determine the activity of these neurons in response to depolarizing and hyperpolarizing synaptic innervation.