To examine the role of circulating fat in the regulation of carbohydrate metabolism, dogs were studied during rest and 90 min of moderate treadmill exercise with nicotinic acid infused to suppress lipolysis with (+Fat; n = 5) or without (-Fat; n = 5) Intralipid. Isotopic and hindlimb arteriovenous methods were used to assess metabolism. Plasma glucose was similar in both protocols during rest and exercise. Differences in insulin, catecholamines, and cortisol between groups were insignificant. Glucagon was approximately 50% greater during rest and exercise in -Fat. The following values represent those at 30 or 40 min of muscular work because peak responses were seen at these times. Arterial free fatty acid levels were 1,129 +/- 253 and 272 +/- 17 mu eq/l at rest and 756 +/- 145 and 269 +/- 51 mu eq/l with exercise in +Fat and -Fat, respectively. Glucose production was 4.2 +/- 0.3 and 5.0 +/- 0.4 mg.kg-1.min-1 at rest and 8.5 +/- 1.3 and 11.4 +/- 0.6 mg.kg-1.min-1 with exercise in +Fat and -Fat, respectively. Glucose utilization was 4.3 +/- 0.3 and 5.3 +/- 0.2 mg.kg-1.min-1 at rest and 9.2 +/- 1.2 and 12.7 +/- 0.8 mg.kg-1.min-1 with exercise in +Fat and -Fat, respectively. Significant glucose flux differences were present during rest and exercise. Limb glucose uptake rose similarly with exercise in +Fat (29 +/- 7 to 82 +/- 22 mumol/min) and -Fat (28 +/- 7 to 88 +/- 16 mumol/min). Arterial blood lactate was 50-100% greater in -Fat compared with that in +Fat.(ABSTRACT TRUNCATED AT 250 WORDS)