Extended Data Fig. 6: Rhythmic activity of DMHPDYN is altered by food access.
a. Schematic of in vivo fiber photometry recording from ARCNPY neurons in mice whose DMHPDYN neurons have been transduced with AAV-FLEX-hM3D-mCherry. b. Effect of DMHPDYN neuron activation on ARCNPY calcium response (n = 5 mice). c. Average ARCNPY activity in squared period (b) after saline (SAL) or DCZ injection (two-tailed paired t-test, *p = 0.028, n = 5 mice). d. In vivo DMHPDYN neuron activities in AL and fasted mice (6 mice). e. Difference in average DMHPDYN neuron activity in dark (ZT14-20) and light (ZT2-8) phases in AL and fasted mice (n = 6 mice, two-tailed paired t-test, **p = 0.005). f. Schematic and representative image of caspase ablation of DMHPDYN neurons (DMHPDYN-X). g. Circadian amplitude of ARCNPY/AGRP activity in intact (n = 5) and DMHPDYN-X mice (n = 3, two-tailed unpaired t-test, *p = 0.013). h. Schematic and representative image of in vivo fiber photometry recording from ARCNPY neurons in mice with chronic DMHPDYN activation. Scale: 200 μm i. Average ARCNPY activity in mice that received DCZ in their drinking water versus control mice (Ctrl: no DCZ, average of 2 days, n = 5 mice each). j. Rayleigh-plot of ARCNPY activity (DCZ mean peak: ZT5.4, Rayleigh p = 0.0123, Ctrl mean peak: ZT6.4, Rayleigh p = 0.0417, phases do not differ significantly). k. Average ARCNPY activity for dark and light phases, two-tailed, paired t-test, *p = 0.012, n = 5 mice. l. Circadian amplitude of ARCNPY activity in control and DCZ groups (n = 5 mice each). m,n. Daily feeding pattern (m, DCZ vs Ctrl *p < 0.05, **p = 0.0036, 5-days average, n = 6 mice each) and total food intake (n, two-tailed, paired t-test, * p = 0.017, 5-days average, n = 6 mice each). o-r. Same as i,k,m,n, but with mice without viral DREADD injection (n = 4 mice each). s. ARCNPY activity in DCZ group fasted at ZT11 (n = 5 mice). t. Rayleigh-plots showing peak time in fasted control and DCZ mice (Control mean peak: ZT17.81, DCZ: ZT4.04, phases differ significantly, p = 0.0154, circular one-way ANOVA). u. Average dark and light time ARCNPY activity in fasted control and DCZ groups (n = 5 mice, two-tailed, unpaired t-test, *p < 0.03, **p = 0.007. Data are presented as mean ± SEM. For exact P-values, and statistics, see Extended Data Fig. 6 Source Data.