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. 2022 Apr 18;134(6):1140–1152. doi: 10.1213/ANE.0000000000006011

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Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the International Anesthesia Research Society.

This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.

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Figure 3. — Chemogenetic stimulation of basal forebrain cholinergic neurons during sevoflurane anesthesia caused electroencephalographic and physiologic activations, increased prefrontal acetylcholine, and produced behavioral arousal. The EEG traces on top are representative frontal EEG segments to show activation in ChAT-Cre rats expressing hM3D(Gq) receptors (left) and the lack of activation in ChAT-Cre rats (control) without hM3D(Gq) receptors (right). Each red (female) and blue (male) dot represents data from 1 rat. Male rats received 1.0 mM C21, while female rats received 0.5 mM C21. Dialysis delivery of compound 21 (C21) into basal forebrain of sevoflurane-anesthetized ChAT-Cre rats expressing hM3D(Gq) receptors significantly increased theta/delta ratio (A) and behavioral arousal score (B). C and E, Respiration rate and heart rate 300 s before (sevoflurane – Sevo) and 300 s after (Sevo + C21) C21-induced EEG changes in ChAT-Cre rats expressing hM3D(Gq) receptors. Dialysis delivery of C21 into basal forebrain significantly increased respiration (C) and heart (E) rates. D and F, C21 delivery into basal forebrain of ChAT-Cre rats expressing only mCherry (control group) failed to elicit significant changes in respiration or heart rate. G and H, Changes in ACh levels in prefrontal cortex (% change from baseline wake state) during Sevo, Sevo + C21, and recovery wake (Rec wake) epochs. The C21 delivery into basal forebrain during sevoflurane anesthesia caused a significant increase in ACh levels to baseline wake levels in ChAT-Cre rats expressing hM3D(Gq) receptors (G) but no such increase was observed in ACh levels in the control group (H). A linear mixed model controlling for sex was used for within-rat statistical comparisons, while a linear regression controlling for sex was used for between-rat (B) statistical comparisons. Post hoc comparisons were Tukey corrected. For B, C, and E: *compared to Control/Sevo. For G and H: *compared to baseline wake state, #compared to Sevo, and §compared to Sevo + C21. The P values are shown at <.05, but the actual P values are reported in the main text and in Supplemental Digital Content 1, Tables S1–S3, http://links.lww.com/AA/D917. A video showing representative behavior after chemogenetic stimulation of basal forebrain cholinergic neurons in ChAT-Cre rats is provided in Supplemental Digital Content 2, Video 1, http://links.lww.com/AA/D918. ACh indicates acetylcholine; EEG, electroencephalogram.