The hyperenergetic-fed beagle dog model of obesity-associated insulin resistance has previously demonstrated lipoprotein abnormalities similar to those of obese insulin-resistant humans. The aim of this study was to check, in the insulin-resistant dog, the mechanism leading to abnormalities in the mass of apolipoprotein B-100 (apo B-100) containing lipoproteins. Six healthy male beagle dogs were overfed with a high-fat diet for 28 +/- 2.5 weeks. Obesity was associated with insulin resistance as assessed by the euglycemic hyperinsulinemic clamp technique. The kinetics of very low-density lipoprotein (VLDL) and low-density lipoprotein (LDL) apo B-100 were recorded in dogs, at healthy and insulin-resistant states, using a primed constant infusion of [5,5,5-D(3)]leucine. Isotopic enrichment was measured by gas chromatography-mass spectrometry (GC-MS). A multicompartmental model was used for the analysis of tracer kinetics data. Apolipoprotein B-100 concentration was higher in VLDL (2.8-fold, P < .05) but lower in LDL (2-fold, P < .05) in the insulin-resistant compared to the healthy state. Kinetic analysis showed a higher VLDL apo B-100 production (1.7-fold, P < .05). The fractional catabolic rate of VLDL did not change significantly, but the lipolysis was decreased significantly (3-fold, P < .05). The lower LDL apo B-100 level in insulin-resistant dogs was explained by a higher LDL fractional catabolic rate (2.5-fold, P < .05). The mechanisms leading to hypertriglyceridemia (higher production rate and lower lipolysis of VLDL) in insulin-resistant dogs were similar to those described in the insulin-resistant humans.