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. 2021 Mar 4;14:628996. doi: 10.3389/fnmol.2021.628996

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Copyright © 2021 Liu, Liu, Zhou, He, Zhang, Cai, Yuan, Luo, Zheng, Yu and Zhang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Chemogenetic manipulation of LHb glutamatergic neurons changed induction and arousal from isoflurane. (A) Chemogenetic activation of LHb glutamatergic neurons shortened loss of righting reflex (LORR) time from 1.4% isoflurane anesthesia. (M3-NS vs M3-CNO, P = 0.000046, paired t-test; Control-CNO vs. hM3Dq-CNO, P = 0.000003, independent-simples t-test; n = 8 per group). (B) Chemogenetic activation of LHb glutamatergic neurons prolonged the recovery of righting reflex (RORR) time from 1.4% isoflurane anesthesia (M3-NS vs M3-CNO, P = 0.000002, paired t-test; EYFP-CNO vs M3-CNO, P = 0.000253, independent-simples t-test; n = 8per group). (C) The power distribution of EEG frequency bands in M3-NS or M3-CNO group under 1.4% isoflurane anesthesia; (δ band: P = 0.000044, paired t-test; α band: P = 0.011, paired t-test). (D) BSR at 20 min before cessation of isoflurane in M3-NS or M3-CNO. BSR is plotted at each minute (n = 8), using two-way analysis of variance (ANOVA) followed by post hoc Bonferroni’s multiple comparisons: F(1, 14) = 8.406, P = 0.0117. (E) Representative EEG waveforms and spectrograms EEG power of M3-CNO and M3-NS under 1.4% isoflurane anesthesia. (F) Chemogenetic inactivation of LHb glutamatergic neurons prolonged the induction time from 1.4% isoflurane anesthesia (M4-NS vs M4-CNO, P = 0.000052, paired t-test; EYFP-CNO vs M4-CNO, P = 0.000002, independent-simples t-test). (G) Chemogenetic inactivation of LHb glutamatergic neurons shortened the recovery time from 1.4% isoflurane anesthesia (M4-NS vs M4-CNO, P = 0.000014, paired t-test; EYFP-CNO vs M4-CNO, P = 0.000003, independent-simples t-test). (H) The power distribution of EEG frequency bands during chemogenetic inactivation of LHb glutamatergic neurons under 1.4% isoflurane anesthesia (M4-NS vs M4-CNO: δ band, P = 0.000312, paired t-test; θ band, P = 0.01, paired t-test; β band, P = 0.01, paired t-test). (I) BSR at 20 min before cessation of isoflurane in M4-NS or M4-CNO. BSR is plotted at each minute (n = 8), using two-way analysis of variance (ANOVA) followed by post hoc Bonferroni’s multiple comparisons: F(1, 14) = 11.66, P = 0.0042. (J) Representative EEG waveforms and spectrograms EEG power of M4-CNO and M4-NS under 1.4% isoflurane anesthesia (*P < 0.05; **P < 0.01; ***P < 0.001; n = 8 per group).