Intracellular free Ca2+ ([Ca2+](i)) is a pivotal signalling element in cell migration and is thought to be required for chemotaxis of Dictyostelium. Ca2+ signalling may also be important for electrotaxis. However this suggestion has been controversial. We show that electric fields direct Dictyostelium cells to migrate cathodally and increase [Ca2+](i) in Dictyostelium cells, as determined by Fluo-3 AM imaging and (45)Ca2+ uptake. Omission of extracellular Ca2+([Ca2+](e)) and incubation with EGTA abolished the electric-field-stimulated [Ca2+](i) rise and directional cell migration. This suggests a requirement for [Ca2+](e) in the electrotactic response. Deletion of iplA, a gene responsible for chemoattractant-induced [Ca2+](i) increase, had only a minor effect on the electric-field-induced [Ca2+](i) rise. Moreover, iplA-null Dictyostelium cells showed the same electrotactic response as wild-type cells. Therefore, iplA-independent Ca2+ influx is necessary for electrotactic cell migration. These results suggest that the [Ca2+](i) regulatory mechanisms induced by electric fields are different from those induced by cAMP and folic acid in Dictyostelium cells. Different roles of the iplA gene in chemoattractant-induced and electrically induced Ca2+ signalling, and different effects of [Ca2+](i) elevation on chemotaxis and electrotaxis indicate that the chemoattractant and electric cues activate distinctive initial signalling elements.