The developmentally regulated expression of prolonged outward potassium currents influences the extent to which sustained inward currents contribute to the action potential at early stages of differentiation. In amphibian spinal neurons, the long duration and calcium dependence of the embryonic action potential and the amount of calcium influx are largely determined by the extent of maturation of the delayed rectifier potassium current (IKv). We have undertaken a parallel study of differentiation of myocytes, in which action potentials are brief and sodium-dependent even at early stages. The early expression of electrical excitability in embryonic amphibian myocytes growing in culture has been examined previously using intracellular voltage recording techniques. The membrane exhibits a delayed rectification in response to depolarization at times earlier than those at which impulses can first be generated. We have examined the differentiation of this outward current in embryonic myocytes developing in vitro, using whole cell voltage clamp. IKv is initially absent. When first recorded it is small and slowly activating but undergoes sixfold increases in both density and rate of activation during the first day in culture. This maturation is dependent upon transcription, and both rate and density are influenced by the presence of other cell types. The large amplitude of the outward delayed rectifier prevents expression of long duration action potentials.