Fig. 2: The neural signature of vigilance state is characterized by low-frequency high-amplitude neuronal synchrony in Dc region of zebrafish.

a Illustration of two-photon calcium imaging of zebrafish dorsal pallium. b Image stack showing divisions of zebrafish dorsal pallium in the HuC:H2B-GCaMP6f fish line. Dc, Dm, Dl, and Dp standing for central, medial, lateral, and posterior zones of dorsal pallium; Vp representing ventral pallium. c Calcium imaging with cellular resolution in Dc region before and after CAS treatment. Each white circle indicates a single neuron. d Analyzed traces of calcium oscillation of 30 neurons in both left and right Dc region as indicated in (c) for 30 min. After CAS treatment, these neurons display synchronized calcium oscillations. e, f Stack image of 6-layer volume-scan of the whole dorsal pallium in zebrafish before and after CAS treatment showing the spatiotemporal pattern of neuronal activity. g Paired Pearson correlation analysis of calcium oscillation between neurons (n = 60) in Dc region before and after CAS treatment, and statistics of correlation coefficient for seven fish. The scale bar indicates Pearson r value. h Continuous decrease of synchronized oscillation numbers is accompanied with a persistent increase of locomotor speed in zebrafish after CAS treatment. N = 5 fish. i Duration of decreased response thresholds to four aversive stimuli and the duration of neuronal synchrony. N = 7 fish. Images in (b, c), e, f represents results from six independent experiments. All data are presented as mean ± SEM. ***P < 0.001; ns denotes no significant difference. For detailed statistics, see Supplementary Table 2. Source data are provided as a Source Data file.