Membrane potentials of the cellular slime mold Dictyostelium discoideum were monitored after chemotactic stimulation by measuring the distribution of the lipophilic cation tetraphenylphosphonium. Stimulation with the chemoattractant cAMP induces a transient membrane hyperpolarization which reaches its most negative value between 1 3 min after stimulation. This hyperpolarization is consistent with the opening of potassium channels. Measurements in streamer F mutant cells reveal that cGMP likely plays a role in the regulation of the cAMP-induced hyperpolarization.