Figure 1

Dopamine phase-shifts PER2::LUC bioluminescence rhythms in mouse RPE-choroid. DA or MLT were added to the culturing media after the third peak of the RPE-choroid PER2::LUC bioluminescence rhythm. The representative data shows application of MLT at CT 14 did not phase-shift the RPE-choroid bioluminescence PER2::LUC rhythm (A). On the other hand, DA application at CT 8 phase-shifted the RPE-choroid bioluminescence rhythm (D). The blue traces indicate controls (vehicle treated) while red traces indicate MLT (A) or DA (D) treated RPE-choroid cultures. The black arrows indicate time of the drug or vehicle (Veh) treatments (A and D). The amount of phase-shift for each individual RPE-choroid rhythm was plotted to create a PRC (B and E). Blue circles indicate cultures treated with Veh and red circles indicate culture treated with either MLT (B) or DA (E). Data were divided into 6 bins at 4-hour intervals for statistical analysis. Data were then used to calculate the phase change of MLT or DA versus their vehicle controls. Bars show the mean amount of phase change from controls and error bars show ±SEM for experimental groups. Error bars from x axis show ±SEM for control groups. MLT did not phase-shift the RPE-choroid PER2::LUC bioluminescence rhythm (n = 4–14 for each bin, Two way ANOVA, p > 0.1, (C) whereas DA significantly phase-shifted PER2::LUC rhythm (Two way ANOVA following Tukey tests, *p < 0.05, **p < 0.01, n = 6–8 for each bin, (F).