05, P = 3.9 × 10−4) and SCN-lesioned (effect of brain area, F3,61 = 2.50, P = 0.068) rats, and they did not differ between the R-MAP and R-Water groups in either SCN-intact rats (interaction between brain area and treatment, F3,60 = 0.91, P = 0.44; main effect of treatment, F1,60 = 3.3 × 10−4, P = 0.99) or SCN-lesioned rats (interaction Dactolisib mw between brain area and treatment, F2,46 = 0.22, P = 0.81; main effect of treatment, F1,46 = 0.21, P = 0.65 for SCN-lesion; Fig. 8B). When compared between the SCN-intact and SCN-lesioned rats, the damping rates did

not differ in either the R-MAP group (interaction between brain area and SCN-lesion, F2,46 = 0.22, P = 0.81; main effect of SCN-lesion, F1,46 = 0.21, P = 0.65) or the R-Water group (interaction between brain area and SCN-lesion, F3,55

= 1.92, P = 0.14; main effect of check details SCN-lesion, F1,55 = 0.95, P = 0.33). The numbers of slices examined were as follows: (i) in the SCN-intact rats: SCN, R-Water, 9; R-MAP, 9; OB, R-Water, 9; R-MAP, 9; CPU, R-Water, 7; R-MAP, 8; PC, R-Water, 5; R-MAP, 1; and SN, R-Water, 9; R-MAP, 8, and (ii) in the SCN-lesioned rats: OB, R-Water, 9; R-MAP, 10; CPU, R-Water, 8; R-MAP, 9; PC, R-Water, 8; R-MAP, 8; and SN, R-Water, 9; R-MAP, 8. The present study clearly demonstrates that restricted MAP drinking at a restricted time of day not only induced MAO in behavior but also entrained it. The free-running of MAO under ad-MAP was modified by the SCN circadian pacemaker entraining to LD. MAO was also expressed in the circadian Per2 rhythms in several extra-SCN brain areas. The Per2 rhythms were phase-shifted by R-MAP. The phase shifts were accelerated by the SCN lesion, especially in the OB and SN, indicating dual regulation of the extra-SCN circadian oscillators in the brain by the SCN and MAO. In the absence of the SCN circadian pacemaker, R-Water also induced circadian oscillation which was not identical with MAO. The oscillatory mechanism underlying MAP-induced behavioral rhythm (i.e., MAO) is suggested as consisting of several extra-SCN oscillators in the brain (Masubuchi et al., 2000) but the exact mechanism is not well understood. A Edoxaban success of ex

vivo analysis of MAO (Natsubori et al., 2013a,b) opened a new experimental approach to this issue, and the fixation of the MAO phase by R-MAP in the present study enabled us to analyse the phase relationships among extra-SCN oscillators in the brain more precisely. The induction of MAO by R-MAP was revealed by subsequent ad-MAP, where the enhanced behavior components at the time of restricted MAP supply showed phase-delay shifts with a period > 24 h. Acceleration and deceleration of phase-delay shifts in MAP-induced behavioral rhythm were observed in the SCN-intact rats but not in the rats with bilateral SCN lesions (Figs 1 and 2). The rate of phase-delay shifts in the SCN-lesioned rats was 1.3 h/day on average and corresponded to a free-running period of 25.3 h.