Activation of the RMTg neurons receiving direct LHb inputs decreases wakefulness.

(A) Left: time course changes of REM sleep and wakefulness of mice in different experimental groups. All mice received LHb injection of AAV2/1–Cre and i.p. injection of CNO (1 mg/kg). EGFP–CNO (n = 6 animals): mice that received RMTg injection of AAV2/9–DIO–EGFP. hM3Dq–CNO (n = 7 animals): mice that received RMTg injection of AAV2/9–DIO–hM3Dq–EGFP. Middle: total REM sleep and wakefulness amounts during the whole day (24 h) of mice in n EGFP–CNO (n = 6 animals) and hM3Dq–CNO groups (n = 7 animals). Right: REM sleep and wakefulness amounts during light phase (ZT 0 –ZT 12) and dark phase (ZT 12 –ZT 24) of mice in n EGFP–CNO (n = 6 animals) and hM3Dq–CNO (n = 7 animals) groups. (B) Number of transitions between different pair of brain states during the light phase (ZT 0 –ZT 12) and dark phase (ZT 12 –ZT 24) of mice in EGFP–CNO (n = 6 animals) and hM3Dq–CNO (n = 7 animals) groups. W–N: Wake to NREM; N–W: NREM to Wake; N–R: NREM to REM. For all figures: one–way ANOVA test, *, P P P (TIF)