Extended Data Figure 10: Activity profiles of flies in which clocks were sped up or slowed down in clock neurons.
From: Circadian clock neurons constantly monitor environmental temperature to set sleep timing
a, b, Sleep plots and sleep slope plots for two Canton S flies (a) and two per01 flies (b). We identified ZT‘A’ (red dashed line) as the local maximum slope value closest to ZT12 (blue dashed line, the onset of cooling) such that sleep slopes at ZT‘A’ and for the next 2 h were non-negative. The difference between these two values (12 − A), which is quantified as the anticipation index, is indicated in each of the plots. See Methods for more details. c, d, To quantify the extent to which the major activity peak anticipates the onset of cooling, we determined the phase of the daily peak of activity closest to ZT12 (the onset of cooling) for individual flies. Representative raw activity profiles of an individual wild-type Canton S fly (c) and per01 mutant fly (d). Red lines represent a smoothed activity profile created by means of a Butterworth low-pass filter (see Methods). The phases of the activity peak closest to the onset of cooling at ZT12 are indicated in the plots. ZT0 corresponds to the start of the heating phase and ZT12 to the start of the cooling phase. e–l, Averaged population activity plots over four days of temperature cycles for the genotypes indicated. The number of flies is shown in m. In all plots, dark lines indicate mean and shaded regions indicate s.e.m. Dashed lines in the plots represent temperature. m, The average phases of the major activity peak closest to the onset of the cooling phase. 0 marks the start of the cooling phase, which corresponds to ZT12 in the locomotor activity plots shown above. The average phase values were calculated by taking the population average of the phase of the major activity peaks of individual flies. Error bars represent s.e.m. Sample sizes are reported to the right of the plot. Negative average phase values indicate that the major activity peak occurred before the onset of the cooling phase, and positive values indicate that the major peak occurred after cooling onset. ANOVAs were conducted to compare Clk4.1m>UAS-DBT(L) flies to Clk4.1m/+ and UAS-DBT(L)/+ controls (F2,95 = 26.1, P < 0.005); Clk4.1m>UAS-DBT(S) flies to Clk4.1m/+ and UAS-DBT(S)/+ controls (F2,76 = 6.5, P < 0.005); Clk856>UAS-DBT(L) flies to Clk856/+ and UAS-DBT(L)/+ controls (F2,78 = 18.4, P < 0.0005); and Clk856>UAS-DBT(S) flies to Clk856/+ and UAS-DBT(S)/+ controls (F2,77 = 25.1, P < 0.0005); Tukey’s tests are indicated. *P < 0.05, ***P < 0.005. Statistical analysis was performed using unpaired two-tailed Student’s t-tests. n, Population averages of the phases of the activity peak closest to the onset of cooling (indicated by the zero line). The number of flies used in the analysis is reported in Extended Data Fig. 6a. Error bars represent s.e.m. An ANOVA was performed for each group of genotypes and Tukey’s honest significant difference tests were conducted for pairwise comparisons. Statistical analysis was performed using unpaired two-tailed Student’s t-tests, **P < 0.005. Individual P values are reported in the Source Data for this figure.
