Figure 1
Representative double-plotted actograms of locomotor activity from Per1-luc transgenic rats fed ad-libitum or under restricted feeding in a 12:12 h light:dark cycle. In the first 5 days all rats were allowed free access to food; one group continued this condition from day 6 to day 15 (A) while another group (B) had a restricted feeding schedule with food available from ZT4 to ZT6 (indicated by the hatched box at the top of the figure inside the bar indicating the L:D cycle). These data are representative of 6 animals.
Figure 2
Per1-luc bioluminescence rhythms in liver explants treated with DMSO. (A) Peak phases (in ZT) from Per1-luc expression rhythms calculated from the first peak occurring at the beginning of the culture. The dashed rectangle inside the subjective light period indicates the former food-restricted schedule, and the vertical dotted line represents the former meal time of the FR rats. (B) Period from Per1-luc expression in liver cultures from AL (left bars) and FR (right bars) rats during and after (washout) DMSO treatment. All data plotted are presented as means ± SEM, n = 6. ++Significant difference between AL vs. FR group on the indicated treatment day (Bonferroni post hoc test, α = 0.05).
Figure 3
Per1-luc bioluminescence rhythms in liver explants treated with BAPTA-AM. (A) Representative daily records of Per1-luc expression in liver explants from AL (superior panel) and FR rats (inferior panel) during BAPTA-AM treatment (100 μM). The grey trace represents the DMSO control and the black trace the BAPTA-AM treated culture. The vertical dotted line represents the former meal time of the FR rats. The horizontal black line above the upper panel indicates the onset and duration of drug treatment which ended with washout of the drug on day 6, marked by the horizontal gray dashed line. The subjective night (or dark phase) is marked only on first day of culture. (B) Period from Per1-luc expression in liver cultures from AL (left bars) and FR (right bars) rats during BAPTA-AM treatment and after washout. All data plotted is presented as means ± SEM, n = 6. *Significant difference vs. the DMSO control and x means difference against washout (Bonferroni post hoc test, α = 0.01).
Figure 4
Per1-luc bioluminescence rhythms in liver explants treated with thapsigargin. (A) Representative daily records of Per1-luc expression in liver explants from AL (superior panel) and FR (inferior panel) rats during thapsigargin treatment (100 μM). The grey trace represents the DMSO control and the black trace the thapsigargin-treated culture. The vertical dotted line represents the former meal time of the FR rats. The horizontal black line above the upper panel indicates the onset and duration of drug treatment which ended with washout of the drug on day 6, marked by the horizontal gray dashed line. The subjective night (or dark phase) is marked only on the first day of culture. (B) Period from Per1-luc expression in liver cultures from AL (left bars) and FR (right bars) rats during thapsigargin treatment and after washout. All data plotted are presented as means ± SEM, n = 6. *Significant difference vs. the DMSO control, and x means difference against washout (Bonferroni post hoc test, α = 0.01).
Figure 5
Per1-luc bioluminescence rhythms in liver explants treated with 2-APB. (A) Representative daily records of Per1-luc expression in liver explants from AL (superior panel) and FR (inferior panel) rats during 2-APB treatment (100 μM). The grey trace represents the DMSO control and the black trace the 2-APB treated culture. The vertical dotted line represents the former meal time of the FR rats. The horizontal black line above the upper panel indicates the onset and duration of drug treatment which ended with washout of the drug on day 6, marked by the horizontal grey dashed line. The subjective night (or dark phase) is marked only on the first day of culture. (B) Period from Per1-luc expression in liver cultures from AL (left bars) and FR (right bars) rats during 2-APB treatment and after washout. All data plotted are presented as means ± SEM, n = 6. *Significant difference vs. the DMSO control, and x means difference against washout (Bonferroni post hoc test, α = 0.01).
Figure 6
Per1-luc bioluminescence rhythms in liver explants treated with ryanodine. (A) Representative daily records of Per1-luc expression in liver explants from AL (superior panel) and FR (inferior panel) rats during ryanodine treatment (100 μM). The grey trace represents the DMSO control and the black trace the ryanodine treated culture. The vertical dotted line represents the former meal time of the FR rats. The horizontal black line above the upper panel indicates the onset and duration of drug treatment which ended with washout of the drug on day 6, marked by the horizontal grey dashed line. The subjective night (or dark phase) is marked only on the first day of culture. (B) Period of Per1-luc expression in liver cultures from AL (left bars) and FR (right bars) rats during ryanodine treatment and after washout. All data plotted are presented as means ± SEM, n = 6. *Significant difference vs. the DMSO control, and x means difference against washout (Bonferroni post hoc test, α = 0.01).