Glucose tolerance is determined by both insulin action and insulin-independent effects, or "glucose effectiveness," which includes glucose-mediated stimulation of glucose uptake (Rd) and suppression of hepatic glucose output (HGO). Despite its importance to tolerance, controversy surrounds accurate assessment of glucose effectiveness. Furthermore, the relative contributions of glucose's actions on Rd and HGO under steady state and dynamic conditions are unclear. We performed hyperglycemic clamps and intravenous glucose tolerance tests in eight normal dogs, and assessed glucose effectiveness by two independent methods. During clamps, glucose was raised to three successive 90-min hyperglycemic plateaus by variable labeled glucose infusion rate; glucose effectiveness (GE) was quantified as the slope of the dose-response relationship between steady state glucose and glucose infusion rate (GE[CLAMP(total)]), Rd (GE[CLAMP(uptake)]) or HGO (GE[CLAMP(HGO)]). During intravenous glucose tolerance tests, tritiated glucose (1.2 microCi/kg) was injected with cold glucose (0.3 g/kg); glucose and tracer dynamics were analyzed using a two-compartment model of glucose kinetics to obtain Rd and HGO components of glucose effectiveness. All experiments were performed during somatostatin inhibition of islet secretion, and basal insulin and glucagon replacement. During clamps, Rd rose from basal (2.54+/-0.20) to 3.95+/-0.54, 6.76+/-1.21, and 9.48+/-1.27 mg/min per kg during stepwise hyperglycemia; conversely, HGO declined to 2.06+/-0.17, 1.17+/-0.19, and 0.52+/-0.33 mg/min per kg. Clamp-based glucose effectiveness was 0.0451+/-0.0061, 0.0337+/-0.0060, and 0.0102+/-0.0009 dl/min per kg for GE[CLAMP(total)], GE[CLAMP(uptake)], and GE[CLAMP(HGO)], respectively. Glucose's action on Rd dominated overall glucose effectiveness (72.2+/-3.3% of total), a result virtually identical to that obtained during intravenous glucose tolerance tests (71.6+/-6.1% of total). Both methods yielded similar estimates of glucose effectiveness. These results provide strong support that glucose effectiveness can be reliably estimated, and that glucose-stimulated Rd is the dominant component during both steady state and dynamic conditions.