Vasomotion, the phenomenon of vessel diameter oscillation, regulates blood flow and resistance. The main parameters implicated in vasomotion are particularly the membrane potential and the cytosolic free calcium in smooth muscle cells. In this study, these parameters were measured in rat perfused-pressurized mesenteric artery segments. The application of norepinephrine (NE) caused rhythmic diameter contractions and membrane potential oscillations (amplitude; 5.3 ± 0.3 mV, frequency; 0.09 ± 0.01 Hz). Verapamil (1 μM) abolished this vasomotion. During vasomotion, 10⁻⁵ M ouabain (Na⁺–K⁺ ATPase inhibitor) decreased the amplitude of the electrical oscillations but not their frequency (amplitude; 3.7 ± 0.3 mV, frequency; 0.08 ± 0.002 Hz). Although a high concentration of ouabain (10⁻³ M) (which exhibits non-specific effects) abolished both electrical membrane potential oscillations and vasomotion, we conclude that the Na+–K+ ATPase could not be implicated in the generation of the membrane potential oscillations. We conclude that in rat perfused-pressurized mesenteric artery, the slow wave membrane type of potential oscillation by rhythmically gating voltage-dependent calcium channels, is responsible for the oscillation of intracellular calcium and thus vasomotion.