Fig. 6: The DN3s integrate DN1as-encoded temperature signals and light signals to modulate heat-induced E peak offset delay and nighttime sleep reduction. | Nature Communications

Fig. 6: The DN3s integrate DN1as-encoded temperature signals and light signals to modulate heat-induced E peak offset delay and nighttime sleep reduction.

From: Dynamic encoding of temperature in the central circadian circuit coordinates physiological activities

Fig. 6: The DN3s integrate DN1as-encoded temperature signals and light signals to modulate heat-induced E peak offset delay and nighttime sleep reduction.

a Schematic of the major upstream neurons of DN3s reconstructed in fly brain template. Left panel is the EM reconstruction of DN3s and upstream neurons. The right panel displays a Sankey histogram depicting the synaptic connections between DN3s and major presynaptic neurons. b, d Representative GCaMP traces (ΔF/F0 ± SEM) of DN3s in response to cooling (b, left, blue, n = 16) and heating (b, right, red, n = 16) in the intact fly (d, black, n = 12) and DN1a-silenced fly (d, red, n = 14). c, e, g Quantification of the relative fold change of calcium activities of DN3s in (b, d, f). Data are presented as mean in ΔF/F0 ± SEM in the histogram; two-tailed t-test; ****p < 0.0001 (c: p = 9.20159E-29; e: p = 5.90626E-16; f: ZT6-8: p = 1.53564E-10; ZT 17-19: p = 3.39847E-05). c related to (b). e related to (d). g related (f). f Representative pseudocolored images of calcium responses of DN3s before and after light pulse (top). Representative GCaMP traces (ΔF/F0 ± SEM) of DN3s in response to light pulse at ZT 6-8 (blue, n = 8) and ZT17-19 (green, n = 9). White arrows indicate DN3s; Scale bars, 50 μm. h, i Representative locomotor activity profiles of control (n = 24), DN3-ablated (h, n = 24) and DN3 optogenetic activation (i, n = 24) flies at 22 °C (gray column) and 30 °C (red column). Activity summed in 30-min bins were plotted. Bar graphs are averages ± SEM; Dark shades indicate lights off. j, k Quantification of E-peak locomotion (column) and E-peak offset delay index (brown plot) in (h, i) at 30 °C (red) and at 22 °C (gray). Data are presented as mean in ΔF/F0 ± SEM; Statistical analysis was conducted using One-Way ANOVA followed by Tukey post-test for multiple comparisons. The letters A, B, C and D above the histograms denote significantly different means within each of the two groups, p < 0.05. Specific p values corresponding to this figure are reported in the Source Data. l A model of DN3s control temperature-induced nighttime wakefulness by receiving ambient temperature information through DN1as. Dark shades indicate lights off (night).

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