We have studied the role of rostral medial prefrontal cortex (mPFC) on reflexively evoked blinks and on classically conditioned eyelid responses in alert-behaving rabbits. The rostral mPFC was identified by its afferent projections from the medial half of the thalamic mediodorsal nuclear complex. Classical conditioning consisted of a delay paradigm using a 370-ms tone as the conditioned stimulus (CS) and a 100-ms air puff directed at the left cornea as the unconditioned stimulus (US). The CS coterminated with the US. Electrical train stimulation of the contralateral rostral mPFC produced a significant inhibition of air-puff-evoked blinks. The same train stimulation of the rostral mPFC presented during the CS-US interval for 10 successive conditioning sessions significantly reduced the generation of conditioned responses (CRs) as compared with values reached by control animals. Interestingly, the percentage of CRs almost reached control values when train stimulation of the rostral mPFC was removed from the fifth conditioning session on. The electrical stimulation of the rostral mPFC in well conditioned animals produced a significant decrease in the percentage of CRs. Moreover, the stimulation of the rostral mPFC was also able to modify the kinematics (latency, amplitude, and velocity) of evoked CRs. These results suggest that the rostral mPFC is a potent inhibitor of reflexively evoked and classically conditioned eyeblinks but that activation prevents only the expression of CRs, not their latent acquisition. Functional and behavioral implications of this inhibitory role of the rostral mPFC are discussed.associative learning ͉ freezing behavior I t is generally accepted that the prefrontal cortex (PFC) represents the highest level in the hierarchical organization of the cortex, and that it is involved in the proper timing, representation, selection, and execution of intentional behaviors and cognitive processes (1-4). Although functions of the PFC cannot be easily ascribed to specific sites, the dorsolateral PFC is more related to working memory tasks and two-interval discrimination (5). In contrast, both medial and orbital PFCs are involved in the emotional component of selected behaviors, including classical and instrumental conditionings (6, 7). In particular, the medial PFC (mPFC) is involved in stimulus salience, sustained attention (8), and/or the integration of learned emotional changes (6), whereas the orbital PFC participates in the selection of appropriate motor actions, exerting an inhibitory control on general motility (1, 9, 10).Although the involvement of the mPFC in associative learning is well documented, the specific roles of its rostral and caudal parts are not yet well established. In this regard, it has been reported that the lesion of the caudal (but not of the rostral) mPFC impairs the acquisition of trace eyeblink conditioning in rabbits (11, 12). The caudal mPFC seems to be necessary at least when weak unconditioned stimulus (US) is used, or during partial reinforcement, large conditioned s...