Low amplitude entrainment of mice and the impact of circadian phase on behavior tests

Abstract

A tremendous increase in the use of genetically engineered mice as experimental animals has led to increased scrutiny of mouse models generally and mouse behavioral paradigms specifically. Although mice are nocturnal, for practical reasons, most experimental procedures, including behavioral studies, are conducted during their inactive, sleep phase. Accumulating evidence indicates that myriad behavioral, cellular and biochemical processes fluctuate with circadian rhythmicity; however, time of day at which experiments are conducted is rarely controlled. The impact of circadian phase on the reliability of experimental results has received little attention and the present data are conflicting. This study addressed two questions. First, will laboratory mice in a typical animal care facility entrain to a low amplitude light cycle using bright/dim rather than light/dark cycles? A positive answer will make reversing photocycle easy to implement in any facility as dim light suitable for animal husbandry and behavioral testing can substitute for darkness during work hours. By monitoring home cage wheel running, we examined the effectiveness of a dim/bright photocycle as a zeitgeiber. We found that mice subjected to dim/bright photocycles effectively entrained such that their subjective night and activity onset coincided with the beginning of the dim light period, suggesting a potential strategy for standardization and management of circadian phase in nocturnal animals. In a second experiment, we asked what effect circadian phase has on behavioral performance in commonly used mouse behavioral tests. We found no main effect of circadian phase on outcome in open field activity, elevated plus maze emotionality, water maze spatial memory, novel object exploration and hyperactivity in response to amphetamine; however, we observed occasional interactions between circadian phase and both strain and sex that were neither consistent nor systematic. These data suggest that the tests examined here are relatively impervious to circadian phase. In general, testing mice during their active phase is more suitable for behavioral studies; a reversed dim/bright photocycle potentially offers one practical strategy for managing rodents' circadian cycles.