选择性光遗传学激活基底前脑和蓝斑核中的食欲素能末梢可促进大鼠从异氟烷麻醉中苏醒。

Selective optogenetic activation of orexinergic terminals in the basal forebrain and locus coeruleus promotes emergence from isoflurane anaesthesia in rats.

机构信息

Department of Anaesthesiology and Perioperative Medicine, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, China.

出版信息

Br J Anaesth. 2021 Jan;126(1):279-292. doi: 10.1016/j.bja.2020.09.037. Epub 2020 Oct 31.

DOI:10.1016/j.bja.2020.09.037

PMID:33131759

Abstract

BACKGROUND

The neuropeptide orexin promotes arousal from general anaesthesia, however the neuronal circuits that mediate this effect have not been defined. We investigated whether orexinergic neurones modulate the basal forebrain (BF) and locus coeruleus (LC) in emergence from anaesthesia.

METHODS

Hcrt rats were generated using a CRISPR/Cas9-based approach. Viruses encoding optogenetic probes were injected into the perifornical lateral hypothalamic (PeFLH) area, optogenetic fibres were embedded in the PeFLH, BF, or LC, and changes in anaesthesia state under 1.4 vol% or 0.8 vol% isoflurane were determined.

RESULTS

In the PeFLH, 98.8% (0.4%) of orexin-A-positive cells expressed tdTomato, and 91.9% (2.2%) of tdTomato cells were orexin-A-positive. Under 1.4 vol% isoflurane anaesthesia, compared with control groups, burst suppression ratio was less, and emergence time was shorter in groups with optogenetic activation of orexinergic cell bodies in the PeFLH (923 [162] vs 493 [68] s, P=0.0003) or orexinergic terminals in the BF (937 (122) vs 674 (108) s, P=0.0049) or LC (913 [128] vs 742 [76] s, P=0.022). Optical stimulation of orexinergic terminals in the BF and LC also improved the movement scores of rats under 0.8 vol% isoflurane anaesthesia.

CONCLUSIONS

Activation of orexinergic terminals in the FB or LC mediates facilitation of emergence from anaesthesia by orexinergic neurones during isoflurane anaesthesia.

摘要

背景

神经肽食欲素可促进全身麻醉后觉醒，但介导该作用的神经元回路尚未确定。我们研究了食欲素能神经元是否调节麻醉苏醒过程中的基底前脑（BF）和蓝斑核（LC）。

方法

采用 CRISPR/Cas9 方法生成 Hcrt 大鼠。将编码光遗传学探针的病毒注入外侧下丘脑穹窿周区（PeFLH），将光遗传纤维埋入 PeFLH、BF 或 LC，在 1.4 体积%或 0.8 体积%异氟醚下测定麻醉状态的变化。

结果

在 PeFLH 中，98.8%（0.4%）的食欲素-A 阳性细胞表达 tdTomato，91.9%（2.2%）的 tdTomato 细胞是食欲素-A 阳性的。在 1.4 体积%异氟醚麻醉下，与对照组相比，PeFLH 食欲素能神经元胞体光遗传学激活组（923[162]与 493[68]秒，P=0.0003）或 BF 食欲素能神经元末梢（937[122]与 674[108]秒，P=0.0049）或 LC（913[128]与 742[76]秒，P=0.022）的爆发抑制比减小，苏醒时间缩短。BF 和 LC 中食欲素能末梢的光刺激也改善了 0.8 体积%异氟醚麻醉下大鼠的运动评分。