Role of GABAergic neuron in bed nucleus of stria terminalis in isoflurane-induced general anesthesia-emergence in mice
10.3760/cma.j.cn131073.20231110.00512
- VernacularTitle:终纹床核GABA能神经元在小鼠异氟烷全麻-觉醒中的作用
- Author:
Xiaoyu GUO
1
;
Huimin WU
;
Dan WANG
;
Hailong DONG
Author Information
1. 徐州医科大学江苏省麻醉学重点实验室 徐州医科大学江苏省麻醉与镇痛应用技术重点实验室 国家药品监督管理局麻醉精神药物研究与评价重点实验室，徐州 221004
- Keywords:
Isoflurane;
Anesthesia, general;
GABAergic neurons;
Bed nucleus of the stria terminalis
- From:
Chinese Journal of Anesthesiology
2024;44(5):587-592
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the role of GABAergic neurons in the bed nucleus of the stria terminalis (BNST) in isoflurane-induced general anesthesia-emergence in mice.Methods:Twenty-three healthy male Vgat-Cre transgenic mice, aged 8-10 weeks, weighing 22 g, were used in the study. In the immunofluorescence staining experiment, 8 mice were selected and divided into 2 groups ( n=4 each) using a random number table method: oxygen group and isoflurane group. Oxygen group inhaled oxygen at a rate of 1.0 L/min for 2 h, while isoflurane group inhaled 1.4% isoflurane + 1.0 L/min oxygen for 2 h. The animals were then sacrificed, and brain tissues were removed and subjected to immunofluorescence staining for determination of the expression of c-Fos and the rate of co-labeling with GABA neurons. For the optogenetic experiment, 15 mice were divided into 3 groups ( n=5 each) using a random number table method: control group (CON group), optogenetic excitation group (CHR2 group) and optogenetic inhibition group (eNpHR group). The rAAV-Ef1a-DIO-mCherry, rAAV-Ef1a-DIO-CHR2-mCherry, and rAAV-Ef1a-DIO-eNpHR3.0-mCherry viruses were injected to the BNST brain region. After 3 weeks of virus expression, the mice were exposed to 1.0% isoflurane + 1.0 L/min oxygen, and their cortical EEG was simultaneously monitored. When the mice reached a stable anesthetic state, optogenetic methods were utilized to modulate the viability of GABAergic neurons in the BNST brain region, and the burst suppression ratio (BSR) of the cortical EEG was recorded at 2 min before light stimulation and 2 min of light stimulation. Results:Compared with oxygen group, the rate of c-Fos co-labeling with GABA neurons in the BNST brain region was significantly reduced ( P<0.05), and the c-Fos-positive neurons were reduced in isoflurane group. Compared with CON group or with the prestimulation level, BSR was significantly decreased in CHR2 group ( P<0.001), and no significant change was found in BSR during light stimulation in eNpHR group ( P>0.05). Conclusions:Decreased viability of GABAergic neurons in the BNST brain region may be involved in the process of loss of consciousness in isoflurane-anesthetized mice, while increased viability of GABAergic neurons in the BNST brain region promotes the transition from anesthesia to emergence.
