Habituation is one of the oldest forms of learning, broadly expressed across sensory systems and taxa. Here, we demonstrate that olfactory habituation induced at different timescales (comprising different odor exposure and intertrial interval durations) is mediated by different neural mechanisms. First, the persistence of habituation memory is greater when mice are habituated on longer timescales. Second, the specificity of the memory (degree of cross-habituation to similar stimuli) also depends on induction timescale. Third, we demonstrate a pharmacological double dissociation between the glutamatergic mechanisms underlying short-and long-timescale odor habituation. LY341495, a class II/III metabotropic glutamate receptor antagonist, blocked habituation only when the induction timescale was short. Conversely, MK-801, an N-methyl-D-aspartate (NMDA) receptor antagonist, prevented habituation only when the timescale was long. Finally, whereas short-timescale odor habituation is mediated within the anterior piriform cortex, infusion of MK-801 into the olfactory bulbs prevented odor habituation only at longer timescales. Thus, we demonstrate two neural mechanisms underlying simple olfactory learning, distinguished by their persistence and specificity, mediated by different olfactory structures and pharmacological effectors, and differentially utilized based solely on the timescale of odor presentation.Habituation, or the progressive reduction in behavioral responsiveness to repeated similar stimuli, is a simple form of learning that underlies animals' capacity to perceptually deemphasize static conditions or inconsequential stimuli in favor of novel, potentially more relevant environmental features or events (Thompson and Spencer 1966;Rose and Rankin 2001). In the laboratory, experimental paradigms based on habituation have been employed to study the properties of nonassociative perceptual learning, to measure the persistence of memory traces, and even to assess the stringency of spontaneous discrimination among similar stimuli. In studies of olfactory spontaneous discrimination using rats or mice, animals habituated to an odorant stimulus demonstrate a cross-habituation response to novel odorant stimuli that are structurally and perceptually similar to the habituated odorant. The degree of cross-habituation declines progressively as test odorants become more dissimilar, and habituation to both the original and cross-habituated odorant stimuli persists for at least several minutes (Cleland et al. 2002;Linster et al. 2002; Mandairon et al. 2006b,c).Olfactory habituation has also been observed in studies in which it is attributed to rapid synaptic adaptation of the mitrapyramidal synapse in the rat anterior piriform cortex (aPC) (Best and Wilson 2004;Yadon and Wilson 2005). Odor-evoked activity in piriform pyramidal neurons adapts within tens of seconds, and this adaptation persists for up to 2 min (Wilson 1998a,b), a substantially faster timescale than that exhibited in the behavioral studies discussed above. Further...