Fig. 4 | Nature Communications

Fig. 4

From: Sleep increases chromosome dynamics to enable reduction of accumulating DNA damage in single neurons

Fig. 4

Neuronal excitation reduces chromosome dynamics in the Rh and SC. ac Simultaneous 2D imaging of neuronal activity (magenta) and chromosome dynamics (green) in the Rh neurons. d Raster plot of ΔF/F of 27 and 26 RCaMP1b-expressing single cells during day and night, respectively. Grayscale: ΔF/F amplitude. e A moderate negative correlation between chromosome dynamics and neuronal activity during day and night (RDay = −0.62, RNight = −0.64, RTotal = −0.64), determined by Pearson correlation coefficient. The average single-cell activity (f, *Pactivity= 0.035) and chromosome dynamics (g, *Pdynamics= 0.03) during day and night. Determined by two-tailed t-test: two samples assuming unequal variance. hl Optogenetic stimulation of neuronal activity reduces chromosome dynamics. hj The neurons expressed both dsRED-TRF1 and ChR2-YFP (arrow) or only dsRED-TRF1 (arrowhead). k, l Chromosome dynamics before and following the light stimuli (ChR2−, n = 9 cells; ChR2+, n = 10 cells). *P = 0.031, determined by two-tailed t-test: two-paired samples for means. Average change is marked by red line. mo Inhibition of neuronal activity during the night does not affect chromosome dynamics. m Representative raster plots of ΔF/F in the Rh of 6 dpf tg(HuC:GCaMP5) larvae under DMSO (n = 6 larvae) or BAPTA-AM (n = 6 larvae) treatments. Grayscale: ΔF/F amplitude. Bottom: histogram of the Ca2+ transients of all cells. n Average spontaneous neuronal activity in single neurons of the Rh under either DMSO (n = 6 larvae) or BAPTA-AM (n = 6 larvae) treatment (ZT23). *P = 2 × 10−4, t = 2.3, degrees of freedom = 7, determined by two-tailed t-test: two samples assuming unequal variance. o Volume of chromosome dynamics during nighttime (ZT23) in DMSO (n = 13 cells) and BAPTA-AM (n = 11 cells)-treated larvae. Values are presented as boxplots and means (black diamonds) or as dot plots and means ± SEM. Boxplots indicate the median and the 25th-to-75th percentiles. The whiskers extend to the most extreme data points. ZT zeitgeber time. Scale bar = 2 µm

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