The inferior colliculus has been implicated in aversive or anxiogenic aspects of defensive behavior. Animals learn to turn off electrical stimulation applied to the inferior colliculus. The purpose of the present study was to determine (1) whether this aversion induced by electrical stimulation can be conditioned to a conditioned stimulus (CS, light) and (2) whether pre-exposure to the CS will diminish the extent of such conditioning, i.e. whether latent inhibition can be established with this paradigm. Rats were placed inside an open field, and thresholds for the escape response to electrical stimulation of the inferior colliculus were determined. The rats were then placed inside a shuttle box and submitted to a two-way avoidance paradigm. Electrical stimulation of the inferior colliculus at the escape threshold was used as negative reinforcement and shuttle box illumination as the CS. The rats quickly learned to avoid or terminate the inferior-colliculus stimulation. Furthermore, the performance of the animals in this paradigm was significantly disrupted when they were pre-exposed to 50 presentations of the CS before the session. These data suggest that the inferior colliculus has neural substrates for supporting associative learning and latent inhibition.