The relationship between acetylcholine (ACh) efflux in medial prefrontal cortex (mPFC) and performance in a visual discrimination task and a variable interval (VI) schedule of reinforcement was studied in rats. Animals were pretrained in one of the two tasks and then unilaterally implanted with microdialysis guide cannula into the mPFC. Animals were then dialyzed, during 12 min collection intervals, in the operant chambers prior to task onset and during and after task performance. Each animal was dialyzed for a total of four sessions: two standard task sessions, one session in which a houselight was flashed at 0.5 Hz during the third 12 min block, and an extinction session (always the last session) in which reinforcement was withheld during the final three blocks. Response accuracy in the discrimination task was very high (> 95% correct) and stable across the four blocks with a progressive increase in omissions. The flashing houselight did not affect performance whereas the loss of reinforcement led to an increase in omissions. VI performance was associated with a high number of lever presses and a high reward rate that declined over the four blocks. Again, the flashing houselight did not affect VI performance whereas lever pressing declined markedly during the extinction session. ACh efflux did not change, relative to baseline, during performance in either task, or with the presentation of the flashing houselight or the loss of reinforcement. These data contrast with the changes in cortical ACh efflux observed in situations characterized by the presentation of novel stimuli or changing demands on attentional processing and, therefore, assist in the specification of hypotheses on the cognitive functions of cortical ACh.