Fig. 7: The DRNGAD2-PVT circuit constrains wakefulness in acute stress condition.
From: A midbrain GABAergic circuit constrains wakefulness in a mouse model of stress
a Schematic of chemogenetic activation of PVT-projecting DRNGAD2 neurons and EEG/EMG recordings. Retro-AAV-FLExloxP-FLP was injected into the PVT and AAV-FLExFRT-hM3Dq-GFP was injected into the DRN of GAD2-Cre mice, respectively. b Representative image showing the expression of hM3Dq-GFP in the DRN. Aq, aqueduct. c EEG power spectrogram, EMG traces, and hypnograms from a DRNGAD2-PVT-hM3Dq mouse during 3 h post saline or CNO (1 mg/kg) injection. Freq., frequency; W, wake; NR: NREM; R: REM. d, Chemogenetic activation of PVT-projecting DRNGAD2 neurons decreases wakefulness. n = 6 mice, two tailed paired t test, wake: t5 = 3.701, P = 0.014; NREM: t5 = 3.48, P = 0.017; REM: t5 = 2.123, P = 0.0872. e Schematic of a 10 min restraint session and following EEG/EMG recordings in DRNGAD2-PVT-hM3Dq mice. f NREM sleep onset latency following a 10 min restraint session. n = 7 mice, two tailed paired t test, t6 = 4.2, P = 0.00569. g Time spent in wake, NREM and REM sleep of DRNGAD2-PVT-hM3Dq mice following restraint. n = 7 mice, two tailed paired t test, wake: t6 = 4.555, P = 0.00387; NREM: t6 = 4.653, P = 0.00349. h Schematic of optogenetic activation of DRNGAD2-PVT circuit. AAV-EF1α-DIO-ChR2-mCherry was injected into the DRN of GAD2-Cre mice and an optic fiber was placed above the PVT. i Heatmaps showing the effects of optogenetic stimulation on tail shock-induced changes of pupil size. j, k Optogenetic activation of DRNGAD2-PVT circuit attenuates tail shock-induced increase of pupil size. n = 6 mice, two tailed paired t test, t5 = 8.63, P = 3.45 × 10−4. Shading in j represents ±SEM. *P < 0.05, **P < 0.01, ***P < 0.001, n.s., not significant. Data (d, f, g, k) are presented as mean ± SEM.
