Volume 28, Issue 13, Pages 2145-2152.e5 (July 2018) Differential Role of Prefrontal and Parietal Cortices in Controlling Level of Consciousness  Dinesh Pal, Jon G. Dean, Tiecheng Liu, Duan Li, Christopher J. Watson, Anthony G. Hudetz, George A. Mashour  Current Biology  Volume 28, Issue 13, Pages 2145-2152.e5 (July 2018) DOI: 10.1016/j.cub.2018.05.025 Copyright © 2018 The Author(s) Terms and Conditions

Figure 1 Cholinergic Stimulation of Prefrontal Prelimbic Cortex in Anesthetized Rat Is Sufficient to Restore Level of Consciousness and Wake-like Behavior (A and B) Behavior after dialysis delivery of 5 mM carbachol (CARB) (A) or 20 mM noradrenaline (NA) (B) into prefrontal prelimbic cortex (PFC) of rats receiving continuous sevoflurane anesthesia (1.9%–2.4%); numbers at the top right corners show the time elapsed (min:s) after CARB/NA reached the target site. (C and D) Demonstration of CARB- (C) and NA-induced (D) electroencephalographic activation. (E–H) Representative spectrograms across the experimental timeline for CARB (E) and NA (G) groups; the vertical color bar is the log scale for power spectral density. Black arrows indicate the approximate time of CARB- and NA-induced electroencephalographic activation quantified as theta/delta ratio in (F) and (H), respectively. (I and J) Effect of CARB on respiration (I) and heart rate (J). (K and L) Effect of NA on respiration (K) and heart rate (L). (M and N) Changes in acetylcholine levels for each rat during sevoflurane anesthesia (Sevo), CARB (M) or NA (N) delivery into PFC during Sevo, and post-Sevo recovery wake (Rec wake) epochs as percent change from pre-anesthesia wake state (Wake). Significance symbols (p < 0.05) show group-level comparisons using Student’s two-tailed paired t test: ∗compared to Wake, #compared to Sevo, and §compared to CARB/NA. Multiple comparisons were Bonferroni corrected. Actual p values are reported in the Results. ACh, acetylcholine; Frtl, frontal; δ, delta; Prtl, parietal; θ, theta; SD, standard deviation. See also Figure S1, Tables S1–S3, and Videos S1 and S2. Current Biology 2018 28, 2145-2152.e5DOI: (10.1016/j.cub.2018.05.025) Copyright © 2018 The Author(s) Terms and Conditions

Figure 2 Cholinergic Stimulation of Posterior Parietal Cortex in Anesthetized Rat Does Not Affect Level of Consciousness (A and B) Behavior after dialysis delivery of 5 mM carbachol (CARB) (A) or 20 mM noradrenaline (NA) (B) into posterior parietal cortex (PPC) of rats receiving continuous sevoflurane anesthesia (1.9%–2.4%); numbers at the top right corners show the time elapsed (min:s) after CARB/NA reached the target site. (C and D) Demonstration of CARB- (C) and NA-induced (D) electroencephalographic activation. (E–H) Representative spectrograms across the experimental timeline for CARB (E) and NA (G) groups; the vertical color bar is the log scale for power spectral density. Black arrows indicate the approximate time of CARB- and NA-induced electroencephalographic activation quantified as theta/delta ratio in (F) and (H), respectively. (I and J) Effect of CARB on respiration (I) and heart rate (J). (K and L) Effect of NA on respiration (K) and heart rate (L). (M and N) Changes in acetylcholine levels for each rat during sevoflurane anesthesia (Sevo), CARB (M) or NA (N) delivery into PPC during Sevo, and post-Sevo recovery wake (Rec wake) epochs as percent change from pre-anesthesia wake state (Wake). Significance symbols (p < 0.05) show group-level comparisons using Student’s two-tailed paired t test: ∗compared to Wake, #compared to Sevo, and §compared to CARB/NA. Multiple comparisons were Bonferroni corrected. Actual p values are reported in the Results. Frtl, frontal; δ, delta; Prtl, parietal; θ, theta; SD, standard deviation. See also Figure S1, Tables S1–S3, and Videos S1 and S2. Current Biology 2018 28, 2145-2152.e5DOI: (10.1016/j.cub.2018.05.025) Copyright © 2018 The Author(s) Terms and Conditions

Figure 3 Cholinergic Stimulation of Medial Parietal Association Cortex in Anesthetized Rat Does Not Affect Level of Consciousness (A) Behavior after dialysis delivery of 5 mM carbachol (CARB) into medial parietal association cortex (MPtA) of rats receiving continuous sevoflurane anesthesia (1.9%–2.4%); numbers at the top right corners show the time elapsed (min:s) after CARB reached the target site. (B) Demonstration of CARB-induced electroencephalographic activation. (C and D) Representative spectrogram across the experimental timeline (C); the vertical color bar is the log scale for power spectral density. Black arrows indicate the approximate time of CARB-induced electroencephalographic activation quantified as theta/delta ratio in (D). (E and F) Effect of CARB delivery on respiration (E) and heart rate (F). (G) Changes in acetylcholine levels for each rat during sevoflurane anesthesia (Sevo), CARB delivery into MPtA during Sevo, and post-Sevo recovery wake (Rec wake) epochs as percent change from pre-anesthesia wake state (Wake). Significance symbols (p < 0.05) show group-level comparisons using Student’s two-tailed paired t test: ∗compared to Wake and #compared to Sevo. Multiple comparisons were Bonferroni corrected. Actual p values are reported in the Results. Frtl, frontal; δ, delta; Prtl, parietal; θ, theta; SD, standard deviation. See also Figure S1 and Tables S1 and S2. Current Biology 2018 28, 2145-2152.e5DOI: (10.1016/j.cub.2018.05.025) Copyright © 2018 The Author(s) Terms and Conditions