Figure 1.
Dynamic activity of VPVglut2 neurons across sleep-wake stages and emotional changes
(A) Schematic of in vivo fiber-photometry recordings of calcium activity of VPVglut2 neurons and EEG/EMGs.
(B) Image showing expression of GCaMP6f in the lateral VP. Dashed box indicates the track of the fiber cannula. Scale bar = 200 μm.
(C) GCaMP6f-expressing cells were also Vglut2 mRNA positive, and quantification of GCaMP6f positive cells (GFP+) double staining for Vglut2 mRNA in the VP. Scale bar = 20 μm. n = 3 mice.
(D) Representative fluorescent traces, heatmap of relative EEG power, and EEG/EMG traces across spontaneous sleep-wake states. The lowest calcium activity was in the NREM sleep, with high EEG delta power and quiet EMG. Calcium activity was higher in wakefulness. The highest calcium activity was in REM sleep.
(E) ΔF/F peaks during wakefulness, NREM sleep, and REM sleep. n = 4 mice, 10 sessions per mouse, ∗∗p < 0.01, one-way ANOVA followed by Tukey’s post hoc test.
(F) Fluorescent signals aligned to arousal state transitions. Upper panel: Individual transitions with color-coded fluorescent intensities (NREM–wake, n = 94; wake–NREM, n = 111; NREM–REM, n = 33; REM–wake, n = 29).
(G) Average calcium transients from all the transitions were expressed as the mean ± SEM. The mean ΔF/F was increased in NREM sleep-to-wake or REM sleep transitions but decreased in wake-to-NREM or REM-to-wake transitions.
(H) Diagram of TS combing calcium recording.
(I–L) Sample traces (I), events (J), above threshold (%) (K), and average peak (%) (L) of calcium activity of VPVglut2 neurons before and after tail suspension. n = 5 mice, ∗p < 0.05, paired t-test.
(M) Heatmap of calcium activity of VPVglut2 neurons in the EPM test. Time ‘0’ indicates transitions from closed-open arms or open-closed arms (30 trials from 5 mice).
(N) Average calcium activity of VPVglut2 neurons in EPM test in closed-to-open arms or open-to-closed arms transitions.
(O) Average ΔF/F in closed and open arms from (N). n = 5 mice, ∗∗p < 0.01, paired t-test.