Specific behavioral associative memory induced by stimulation of the cortically-projecting cholinergic nucleus basalis (NB) is dependent on intrinsic acetylcholine and shares with natural memory such features as associativity, specificity, rapid formation, consolidation and long-term retention. Herein, we examined extinction and the effects of stimulus pre-exposure. Two groups of adult male rats (n=4 each) were first tested for behavioral responses (disruption of ongoing respiration) to tones (1-15 kHz), constituting a pre-training behavioral frequency generalization gradient (BFGG). They next received a first session of training, 200 trials of a tone (8.00 kHz, 70 dB, 2 s) either paired with electrical stimulation of the NB (100 Hz, 0.2 s, approximately 67 microA, NBstm) (group IP) or unpaired (group IU). Twenty-four hours later, they were tested for behavioral memory by obtaining post-training BFGGs. Then the contingencies were reversed yet another 24 h later; the IP group received tone and NBstm unpaired and the IU group received them paired. A final set of generalization gradients was obtained the next day. All stimuli were presented with subjects under state control indexed by regular respiration. Tested 24 h post-initial training, the IP group developed specific associative behavioral memory indicated by increased responses only to CS-band frequencies, while the IU group did not. After subsequent training with unpaired stimuli, the IP group exhibited experimental extinction. Furthermore, after initial exposure to the CS and NBstm unpaired, the IU group exhibited a tendency toward reduced conditioning to CS/NBstm pairing and a significant increase in latency of conditioned responses. The present findings provide additional support for the hypothesis that engagement of the NB is sufficient to induce natural associative memory and suggest that activation of the NB may be a normal component in the formation of natural associative memory.