We examined the effects of endurance training on parameters of glucose flux during rest and exercise in postmenopausal women. Ten sedentary, but healthy women (55 ± 1 yr) subjects completed 12 weeks of endurance exercise training on a cycle ergometer [5 days per week, 1 h per day, 65% peak oxygen consumption (VO₂peak)]. Flux rates were determined by primed-continuous infusion of [6,6-²H]glucose (D₂-Glucose) during 90 minutes of rest and 60 min of cycle ergometer exercise during one pre-training exercise trial [65% VO₂peak (PRE)] and two post-training exercise trials [the power output that elicited 65% pre-training VO₂peak (ABT) and 65% post-training VO₂peak (RLT)]. Training increased VO₂peak by 16.3 ± 3.9% (p < 0.05). Epinephrine and glucagon were lower during ABT and lactate was lower during ABT and RLT (p < 0.05), but the apparent insulin response was unchanged. Whole body glucose Ra decreased post-training during exercise at a given power output (4.58 ± 0.39 mg kg^-1 min^-1 during ABT compared to 5.21 ± 0.48 mg kg^-1 min^-1 PRE, p < 0.05), but not at the same relative workload (5.85 ± 0.36 mg kg^-1 min^-1). Training resulted in a 35% increase in glucose MCR during exercise at the same relative intensity (7.16 ± 0.42 ml kg^-1 min^-1 during RLT compared to 5.28 ± 0.42 ml kg^-1 min^-1 PRE, p < 0.05). Changes in parameters of glucose kinetics during exercise were accomplished without changes in dietary composition, body weight or body composition. We conclude that despite changes in the hormonal milieu that occurs at menopause, endurance training results in a similar magnitude in training-induced alterations of glucose flux as seen previously in younger women.