Wing high dose AM281 during the first 3 Hr of the DP (ZT12-15: t (191.99) = 3.15, p = 0.004), which was the same point in the circadian cycle when NREM sleep time was increased in the experiment where AM281 was delivered before the DP. For NREM bout duration, there was an overall interaction (treatment x time of day within photoperiod, F (18, 141.98) = 4.74, p < 0.001), a secondary interaction (treatment x photoperiod, F(2, 82.68) = 12.61, p < 0.001), and main effects of both treatment (F(2, 59.88) = 9.86, p < 0.001) and Necrostatin-1 site photoperiod (F(1, 109.96) = 31.83, p < 0.001). High dose AM281 substantially reduced NREM bout duration (t(62.07) = -4.42, p < 0.001), particularly during the LP (t(69.68) = -6.23, p < 0.001). More specifically, NREM bout duration was reduced for the first 3 Hr of the LP following low dose AM281 (ZT00-03: t(176.12) = -2.82, p = 0.011) and for the first 9 Hr following high dose AM281 (ZT00-09: t(176.12) -3.46, p 0.001). The number of NREM bouts was affected in the opposite manner. There was an overall interaction (treatment x time of day within photoperiod, F(18, 144.093) = 3.266, p < 0.001), a secondary interaction (treatment x photoperiod, F (2, 78.77) = 13.65, p < 0.001), and main effects of both treatment (F(2, 53.14) = 19.99, p < 0.001) and photoperiod (F(1, 113.94) = 148.145, p < 0.001). High dose AM281 increased the number of NREM bouts (t(63.61) = 6.79, p < 0.001) particularly during the first 9 Hr of the LP (ZT00-09: t(159.32) ! 4.22, p < 0.001), and both doses increased the number of NREM bouts during the second quarter of the DP (ZT15-18: t(159.32) ! 2.31, p 0.045). Thus, blockade of CB1 receptors greatly fragments NREM sleep, but opposing effects on NREM bout duration and the number of NREM bouts result in subtle changes in scan/nsw074 total sleep time. In addition to the effects on NREM sleep, REM sleep time was significantly reduced following AM281 administration prior to the LP (Fig 10C, BX795 chemical information bottom row). For the percent of time spent in REM sleep, there was a secondary interaction (treatment x photoperiod, F(2, 95.51) = 36.30, p < 0.001), nested interaction (time of day within photoperiod, F(6, 143.275) = 11.15, p < 0.001), and main effects of both treatment (F(2, 74.2) = 21.38, p < 0.001) and photoperiod (F(1, 116.42) = 68.40, p < 0.001). Both low and high dose AM281 reduced REM sleep during the jir.2012.0140 LP (t(84.01) -4.22, p < 0.001). Low dose AM281 reduced REM sleep during the first 6 Hr of the LP (ZT00-06: t(190.28) -3.30, p 0.002), and high dose AM281 reduced REM at all times during the LP (ZT00-12: t(190.28) -3.40, p 0.002). For REM bout duration, there was a secondary interaction (treatment x photoperiod, F(2, 81.71) = 7.10, p = 0.001), nested interaction (time of day within photoperiod, F(6, 139.18) = 5.34, p < 0.001), and a main effect of treatment (F(2, 56.07) = 3.35, p = 0.042). The high dose of AM281 reduced REM bout duration during the LP (t(64.52) = -4.05, p < 0.001), particularly during the first 9 Hr of the LP (ZT00-09: t(156.05) -2.31, p 0.044). For the number of REM bouts, there was an overall interaction (treatment x time of day within photoperiod, F(12, 133.54) = 1.85, p = 0.046), secondary interaction (treatment x photoperiod, F(2, 68.89) 7.463, p = 0.001), nested interaction (time of day within photoperiod, F(6, 127.15) = 3.99, p = 0.001), and main effects of both treatment (F(2, 46.46) = 6.39, p = 0.004) and photoperiod (F(1, 99.89) = 59.86, p < 0.001). The number of REM bouts were reduced by high dose AM281.Wing high dose AM281 during the first 3 Hr of the DP (ZT12-15: t (191.99) = 3.15, p = 0.004), which was the same point in the circadian cycle when NREM sleep time was increased in the experiment where AM281 was delivered before the DP. For NREM bout duration, there was an overall interaction (treatment x time of day within photoperiod, F (18, 141.98) = 4.74, p < 0.001), a secondary interaction (treatment x photoperiod, F(2, 82.68) = 12.61, p < 0.001), and main effects of both treatment (F(2, 59.88) = 9.86, p < 0.001) and photoperiod (F(1, 109.96) = 31.83, p < 0.001). High dose AM281 substantially reduced NREM bout duration (t(62.07) = -4.42, p < 0.001), particularly during the LP (t(69.68) = -6.23, p < 0.001). More specifically, NREM bout duration was reduced for the first 3 Hr of the LP following low dose AM281 (ZT00-03: t(176.12) = -2.82, p = 0.011) and for the first 9 Hr following high dose AM281 (ZT00-09: t(176.12) -3.46, p 0.001). The number of NREM bouts was affected in the opposite manner. There was an overall interaction (treatment x time of day within photoperiod, F(18, 144.093) = 3.266, p < 0.001), a secondary interaction (treatment x photoperiod, F (2, 78.77) = 13.65, p < 0.001), and main effects of both treatment (F(2, 53.14) = 19.99, p < 0.001) and photoperiod (F(1, 113.94) = 148.145, p < 0.001). High dose AM281 increased the number of NREM bouts (t(63.61) = 6.79, p < 0.001) particularly during the first 9 Hr of the LP (ZT00-09: t(159.32) ! 4.22, p < 0.001), and both doses increased the number of NREM bouts during the second quarter of the DP (ZT15-18: t(159.32) ! 2.31, p 0.045). Thus, blockade of CB1 receptors greatly fragments NREM sleep, but opposing effects on NREM bout duration and the number of NREM bouts result in subtle changes in scan/nsw074 total sleep time. In addition to the effects on NREM sleep, REM sleep time was significantly reduced following AM281 administration prior to the LP (Fig 10C, bottom row). For the percent of time spent in REM sleep, there was a secondary interaction (treatment x photoperiod, F(2, 95.51) = 36.30, p < 0.001), nested interaction (time of day within photoperiod, F(6, 143.275) = 11.15, p < 0.001), and main effects of both treatment (F(2, 74.2) = 21.38, p < 0.001) and photoperiod (F(1, 116.42) = 68.40, p < 0.001). Both low and high dose AM281 reduced REM sleep during the jir.2012.0140 LP (t(84.01) -4.22, p < 0.001). Low dose AM281 reduced REM sleep during the first 6 Hr of the LP (ZT00-06: t(190.28) -3.30, p 0.002), and high dose AM281 reduced REM at all times during the LP (ZT00-12: t(190.28) -3.40, p 0.002). For REM bout duration, there was a secondary interaction (treatment x photoperiod, F(2, 81.71) = 7.10, p = 0.001), nested interaction (time of day within photoperiod, F(6, 139.18) = 5.34, p < 0.001), and a main effect of treatment (F(2, 56.07) = 3.35, p = 0.042). The high dose of AM281 reduced REM bout duration during the LP (t(64.52) = -4.05, p < 0.001), particularly during the first 9 Hr of the LP (ZT00-09: t(156.05) -2.31, p 0.044). For the number of REM bouts, there was an overall interaction (treatment x time of day within photoperiod, F(12, 133.54) = 1.85, p = 0.046), secondary interaction (treatment x photoperiod, F(2, 68.89) 7.463, p = 0.001), nested interaction (time of day within photoperiod, F(6, 127.15) = 3.99, p = 0.001), and main effects of both treatment (F(2, 46.46) = 6.39, p = 0.004) and photoperiod (F(1, 99.89) = 59.86, p < 0.001). The number of REM bouts were reduced by high dose AM281.
