Exposure of mice to a 24 h light:dark:light:dark (LDLD) cycle with dimly illuminated nights induces the circadian timing system to program two intervals of activity and two intervals of rest per 24 h cycle and subsequently allows entrainment to a variety of extraordinary light regimens including 30 h LDLD cycles. Little is known about critical lighting requirements to induce and maintain this non-standard entrainment pattern, termed “bifurcation,” and to enhance the range of apparent entrainment. The current study determined the necessary duration of the photophase for animals to bifurcate and assessed whether requirements for maintenance differed from those for induction. An objective index of bifurcated entrainment varied with length of the photophase over 4–10 h durations, with highest values at 8 h. To assess photic requirements for the maintenance of bifurcation, mice from each group were subsequently exposed to the LDLD cycle with 4 h photophases. While insufficient to induce bifurcation, this photoperiod maintained bifurcation in mice transferred from inductive LDLD cycles. Entrainment to 30 h LDLD cycles also varied with photoperiod duration. These studies characterize non-invasive tools that reveal latent flexibility in the circadian control of rest/activity cycles with important translational potential for addressing needs of human shift-workers.
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