Glucagon was infused to maintain plasma concentrations three to six times the basal level (300 to 600 pg per milliliter) into 16 normal and seven non-diabetic obese subjects. Hyperglucagonemia caused only a transient rise of 5 to 10 mg per 100 ml in basal glucose levels and had no effect on oral glucose tolerance or plasma insulin. In three patients with adult and two with juvenile-onset diabetes on maintenance insulin, hyperglucagonemia maintained for two to four days caused no change in plasma glucose of ketone concentration. In contrast, in nine insulin-withdrawn patients the glycemic response to hyperglucagonemia was five to 15 times greater (P less then 0.05) than in normal controls. Hyperglucagonemia does not cause glucose intolerance in normal subjects or bring about deterioration of diabetic control when insulin is available. Glucagon in the insulin-deprived patient can worsen the diabetic state. These findings suggest the primary role of insulin deficiency in the diabetogenic action of glucagon.