Glucose tolerance was measured in (nocturnal) mice exposed to light-dark stimulus patterns simulating those that (diurnal) humans would experience while working dayshift (DSS) and 2 rotating night shift patterns (1 rotating night shift per week [RSS1] and 3 rotating night shifts per week [RSS3]). Oral glucose tolerance tests were administered at the same time and light phase during the third week of each experimental session. In contrast to the RSS1 and RSS3 conditions, glucose levels reduced more quickly for the DSS condition. Glucose area-under-The-curve measured for the DSS condition was also significantly less than that for the RSS1 and RSS3 conditions. Circadian disruption for the 3 light-dark patterns was quantified using phasor magnitude based on the 24-h light-dark patterns and their associated activity-rest patterns. Circadian disruption for mice in the DSS condition was significantly less than that for the RSS1 and RSS3 conditions. This study extends previous studies showing that even 1 night of shift work decreases glucose tolerance and that circadian disruption is linked to glucose tolerance in mice.