Evidence suggests that excitatory amino acid-containing afferents from the prefrontal cortex (PFC) play an important role in the induction of burst firing in midbrain dopaminergic (DA) neurons. In the present study, the extracellular activity of individual DA neurons (A10 and A9 cell groups) was recorded during single pulse electrical stimulation (0.25 and 1 mA) of the PFC. The majority of cells were responsive, and two main patterns of activity were elicited: responses characterised by an initial excitation (E responses; 41.8% of responses at 0.25 mA and 26.6% at 1 mA; cell groups combined) and responses characterised by excitation following an initial inhibition (IE responses; 43.3% of responses at 0.25 mA and 56.6% at 1 mA; cell groups combined). Burst analysis performed on the excitatory phase of E and IE responses revealed that the excitation contained events which fulfilled the criteria for natural bursts in DA neurons. A procedure was developed for assessing whether these bursts were time-locked to the stimulus. This showed that 27.9% of E responses and 33.3% of IE responses were accompanied by time-locked bursts (currents and cell groups combined). It is argued that time-locked bursts during IE responses were produced by rebound activation of a low threshold calcium conductance, whereas time-locked bursts during E responses were produced by excitatory afferents. Since natural bursts in DA neurons also seem to involve cortically induced excitation, the hypothesis that the PFC plays a role in the production of natural bursts in DA neurons is strengthened.