(A) Schematic illustration of the photo-inhibition paradigm
during NREMs and REMs in MCH-Cre mice. Yellow/orange light (593.5 nm wave
length) illuminations were applied after 30 seconds of stable NREMs and
continued until the next wake bout irrespective of whether NREMs transitioned
into REMs or wake. (B) Percentage of NREMs bouts transitioning to
REMs. Repeated measures (RM) two-way ANOVA for ‘Drug treatment’
(F(1,8)=32.95, p=0.0004) and ‘Laser treatment’
(F(1,8)=49.73, p=0.0001), followed by Holm-Sidak’s
multiple comparison test (saline + sham inhibition vs.
saline + laser light, p=0.0068; saline + sham
inhibition. vs. CNO + sham inhibition, p=0.0363;
saline + sham inhibition vs. CNO + laser
light, p=0.0309; saline + laser light vs.
CNO + sham inhibition, p=0.0004; CNO + sham
inhibition vs. CNO + laser light, p=0.0012). RM two-way ANOVAs for
the duration of REMs bouts (C) and the latency to REMs onset
(D) were not statistically significant (REMs bout duration
‘Drug treatment’ F(1,8)=2.853, p=0.1297, ‘Laser
treatment’ F(1,8)=0.452, p=0.5203; REMs latency ‘Drug
treatment’ F(1,8)=1.797, p=0.2169, ‘Laser
treatment’ F(1,8)=0.5166, p=0.4928). (E) NREMs
bout duration. RM two-way ANOVA for ‘Drug treatment’
(F(1,8)=8.766, p=0.0181) and ‘Laser treatment’
F(1,8)=19.3, p=0.0023, followed by Holm-Sidak’s multiple
comparison test (saline + sham inhibition. vs. CNO +
laser light, p=0.0282). (F) Percentage of NREMs during
the 15 trials (starting with NREMs) within the 3h recording period. RM two-way
ANOVA for ‘Drug treatment’ (F(1,8)=6.879, p=0.0305) and
‘Laser treatment’ (F(1,8)=61.61, p=0.≤0.0001),
followed by Holm-Sidak’s multiple comparisons test
(saline + sham inhibition vs.
saline + laser light, p=0.05;
saline + laser light vs.
CNO + sham inhibition, p=0.0127;
CNO + sham inhibition vs.
CNO + laser light, p=0.0346).
(G) Percentage of REMs during the 15 trials (starting with
NREMs) within the 3h recording period. RM two-way ANOVA for ‘Drug
treatment’ (F(1,8)=6.879, p=0.0305) and ‘Laser
treatment’ (F(1,8)=61.61, p≤0.0001), followed by
Holm-Sidak’s multiple comparisons test (saline +
sham inhibition vs. saline + laser
light, p=0.05; saline + laser
light vs. CNO + sham inhibition,
p=0.0127; CNO + sham inhibition vs.
CNO + laser light, p=0.0346). Saline or
CNO (0.3 mg/kg; i.p.) was injected at ZT6 and sham/laser light illumination was
applied between ZT7-ZT10 (n = 9 mice). Data are Mean ± SEM, * P <
0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Scale
bar in (A) is 20 sec.