The locations of the vascular resistance sites which regulate vascular tone in the hepatic arterial and portal venous vasculatures of the rat liver were identified using a new, in vitro, dual-perfused liver preparation. Twelve livers of male Wistar rats were perfused via the hepatic artery and portal vein at fixed flow and at physiological pressure. Dose-related vasoconstriction to injections or infusions of noradrenaline was measured as transient or sustained increases in perfusion pressure, respectively, in the hepatic arterial and portal venous vasculatures. Direct injections/infusions of noradrenaline refer to those administered into the vasculature from which pressure was recorded, e.g., the effects of hepatic arterial (direct) injections/infusions of noradrenaline upon hepatic arterial perfusion pressure. Indirect injections/infusions of noradrenaline were those administered to the adjacent afferent vasculature, e.g., the effects of portal venous (indirect) injections of noradrenaline upon hepatic arterial perfusion pressure. The converse applies for recordings of portal venous perfusion pressure. The −log( M) ED 50 values to direct (hepatic arterial) and indirect (portal venous) injections in the hepatic artery were 4.25±0.20 and 3.40±0.10, respectively, and were significantly different ( P<0.01, Student's unpaired t-test); the −log( M) ED 50 values to direct (portal venous) and indirect (hepatic arterial) injections in the portal vein were 3.91±0.08 and 3.85±0.11, respectively, and were not significantly different ( P>0.05, Student's unpaired t-test). Similarly, the −log( M) ED 50 values to direct (hepatic arterial) and indirect (portal venous) infusions in the hepatic artery were 5.28±0.11 and 3.75±0.12, respectively, and were significantly different ( P<0.01, Student's unpaired t-test); the −log( M) ED 50 values to direct (portal venous) and indirect (hepatic arterial) infusions in the portal vein were 5.31±0.19 and 5.70±0.16, respectively, and were not significantly different ( P>0.05, Student's unpaired t-test). These results demonstrated that there is little transfer of noradrenaline from the portal venous to the hepatic arterial resistance sites, but significant transfer from the hepatic artery to the portal venous suggesting that; (a) the portal venous resistance sites are located at the sinusoidal or post-sinusoidal level; and (b) the hepatic arterial resistance sites are located at the pre-sinusoidal level.