Potassium channels have been implicated in central roles in activity-dependent neural plasticity. The giant fiber escape pathway ofDrosophilahas been established as a model for analyzing habituation and its modification by memory mutations in an identified circuit. Several genes inDrosophilaencoding K+channel subunits have been characterized, permitting examination of the contributions of specific channel subunits to simple conditioning in an identified circuit that is amenable to genetic analysis. Our results show that mutations altering each of four K+channel subunits (Sh,slo,eag, andHk) have distinct effects on habituation at least as strong as those ofdunceandrutabaga, memory mutants with defective cAMP metabolism (Engel and Wu, 1996). Habituation, spontaneous recovery, and dishabituation of the electrically stimulated long-latency giant fiber pathway response were shown in each mutant type. Mutations ofSh(voltage-gated) andslo(Ca2+-gated) subunits enhanced and slowed habituation, respectively. However, mutations ofeagandHksubunits, which confer K+-current modulation, had even more extreme phenotypes, again enhancing and slowing habituation, respectively. In double mutants,Shmutations moderated the strong phenotypes ofeagandHk, suggesting that their modulatory functions are best expressed in the presence of intactShsubunits. Nonactivity-dependent responses (refractory period and latency) at two stages of the circuit were altered only in some mutants and do not account for modifications of habituation. Furthermore, failures of the long-latency response during habituation, which normally occur in labile connections in the brain, could be induced in the thoracic circuit stage inHkmutants. Our work indicates that different K+channel subunits play distinct roles in activity-dependent neural plasticity and thus can be incorporated along with second messenger “memory” loci to enrich the genetic analysis of learning and memory.