Sleep restriction leads to impairments in glucose regulation in healthy individuals. Exercise can improve glucose regulation and hence may be beneficial for attenuating the impairment in glucose regulation after sleep restriction. This study aimed to investigate whether an acute bout of exercise performed the morning after a single night of sleep restriction would improve glucose regulation compared to no exercise. Twelve healthy males were recruited for this randomised crossover study which consisted of four conditions – control (CON), sleep restriction (SR), control plus exercise (CON+Ex), and sleep restriction plus exercise (SR+Ex). Time in bed was 8 hr in the control conditions (approximately 23:00 – 07:00) and 4 hr in the sleep restriction conditions (approximately 03:00 – 07:00). Participants underwent a 1 week entraining period prior to each condition, during which they kept a consistent bed and wake time. Compliance was assessed by wrist actigraphy throughout the study. After consuming a 24 hr controlled diet participants arrived at the laboratory at 08:00 in a fasted state and conducted either a high intensity exercise bout or rested for the equivalent duration. The exercise protocol consisted of 4 all-out 30-s sprints against 7.5% of bodyweight, separated by 4.5 min of active recovery. Following a 30 min rest period an oral glucose tolerance test was conducted. Seven blood samples were obtained over a 2 hour period. Serum glucose and insulin were analysed from the blood samples. Total (0-120 min), early phase (0-60 min) and late phase (60-120 min) area under the curve (AUC) for glucose and insulin were calculated using the trapezoidal rule. Matsuda index and HOMA-IR were used to estimate whole-body insulin sensitivity. Repeated measures ANOVAs were used to determine differences between conditions. Total AUC was not significantly different between conditions for glucose (P=0.261) or insulin (P=0.678). Similarly, early and late phase glucose AUC and early phase insulin AUC did not differ between conditions (P>0.05 for all). However, late phase insulin AUC showed a significant main effect (P=0.010). Post-hoc analysis revealed a significantly lower late phase AUC in SR+Ex than SR (P=0.025). Matsuda index and HOMA-IR showed a main effect of condition (P=0.029 and P=0.009, respectively), with a trend towards increased HOMA-IR in SR compared to CON (P=0.053). These findings suggest that exercise improves the late post-prandial phase following a single night of sleep restriction. Whilst no effect of exercise on the Matsuda index or HOMA were found, the lower late-phase insulin AUC following exercise suggests that there may be some benefit of performing a single bout of high intensity exercise following a night of sleep restriction. This may have implications for individuals who face unplanned sleep curtailment.