This study was undertaken to determine whether the increase in plasma glucagon concentration that occurs in response to prolonged exercise is modified by endurance exercise training. Eight subjects participated in an exercise program, consisting of running and bicycling, 4 days/wk for 10 wk. The training program resulted in an average increase in VO2 max of 18%. The average increase in plasma glucagon during a 60-min long bicycle exercise test that required 60% of the subjects' VO2 max was 107+/-28 pg/ml, from 116+/-14 pg/ml at rest to 223+/-37 pg/ml after 60 min of exercise, prior to training. After training the same absolute work rate resulted in an increase in plasma glucagon of only 20+/-6 pg/ml, from 125+/-20 to 145+/-16 pg/ml (P less than 0.02). A similar blunting of the glucagon response to exercise was seen during work of the same relative intensity after training. Plasma insulin concentration decreased from 18.1+/-2.5 to 7.6+/-1.6 muunits/ml during the 60 min of exercise before training. A similar decrease in insulin concentration was seen at the same relative work rate after training. However, the decrease in plasma insulin at the same absolute work rate, from 18.5+/-3.0 to 12.5+/-1.8 muunits/ml, was significantly smaller after training (P less than 0.05).