The aim of this study was to examine the influence of small changes in glucagon on hepatic glucose production during a euglycemic, hyperinsulinemic clamp. During 1.0 mU/kg · min insulin infusion, euglycemia was maintained by glucose infusion and glucagon was infused at various rates so as to cause plasma glucagon levels to increase, decrease, or remain unchanged. Changes in glucagon were found to be positively associated with changes in glucose production and inversely related to the degree of suppression of tracer or arteriovenous difference determined endogenous glucose production. Thus, animals in which the glucagon levels increased, appeared to have decreased hepatic insulin sensitivity, while animals in which glucagon levels decreased, appeared to have increased insulin sensitivity. In conclusion, since glucagon often declines during a euglycemic hyperinsulinemic clamp, and since small changes in glucagon can have marked effects on the suppression of hepatic glucose output even in the presence of high insulin levels, changes in glucagon should be considered when conclusions regarding hepatic insulin sensitivity are being drawn.