异氟烷麻醉小鼠皮质2/3层神经活动的变化:高分辨光场智能成像显微平台观测  

Changes in activity of layer 2/3 cortical neurons in isoflurane-anesthetized mice:Real-time Ultra-large-Scale High-resolution imaging platform

作　　者：王紫霖 王鸣瑞 庄超玮 张汝金 陈昆莎 曹江北[1] 米卫东[1] Wang Zilin;Wang Mingrui;Zhuang Chaowei;Zhang Rujin;Chen Kunsha;Cao Jiangbei;Mi Weidong(Department of Anesthesiology,the First Medical Center,Chinese PLA General Hospital,Beijing 100853,China;Tsinghua-Berkeley Shenzhen Institute,Tsinghua University,Shenzhen 518055,China;Department of Automation,Tsinghua University,Beijing 100084,China)

机构地区：[1]解放军总医院第一医学中心麻醉科,北京100853 [2]清华大学清华伯克利深圳学院,深圳518055 [3]清华大学自动化系,北京100084

出　　处：《中华麻醉学杂志》2022年第10期1202-1206,共5页Chinese Journal of Anesthesiology

摘　　要：目的采用高分辨光场智能成像显微平台对异氟烷麻醉小鼠皮质2/3层神经活动进行观测并分析。方法清洁级健康雄性Rasgrf2-Cre/Ai148d小鼠,8~12周龄,体重18~25 g。行颅骨替代手术后,恢复10 d进行下一步实验。采用高分辨光场智能成像显微平台对头部固定的小鼠皮质2/3层神经元钙荧光信号进行成像数据采集。清醒状态、1.2%异氟烷麻醉和麻醉结束后的数据采集时间分别为100、600和600 s。成像数据采用Image J软件以及MATLAB软件进行分析。结果小鼠皮质2/3层神经活动整体趋势随1.2%异氟烷吸入后先下降,随后趋于稳定;异氟烷停止吸入后,小鼠皮质神经活动逐渐增强。停止吸入异氟烷后,不同脑区神经活动恢复速率存在差异。与其他脑区相比,运动皮质神经活动存在恢复延迟。麻醉维持期间,小鼠压后皮质2/3层大部分神经元活动减弱,但部分神经元活动增强。结论在整体观测层面、脑区层面以及单细胞层面,异氟烷麻醉小鼠皮质2/3层神经活动具有非一致性,提示麻醉诱导和苏醒过程由不同的神经通路参与。Objective To observe and analyze the changes in activity of layer 2/3 cortical neurons in isoflurane-anesthetized mice by Real-time Ultra-large-Scale High-resolution(RUSH)imaging platform.Methods Clean-grade healthy male Rasgrf2-Cre/Ai148d mice,aged 8-12 weeks,weighing 18-25 g,were studied.The mice recovered ten days after the skull replacement surgery and proceeded to the next experiment.Imaging data of calcium fluorescence signals from layer 2/3 cortical neurons were acquired by RUSH imaging platform after fixing the head of mice.The time of imaging data acquisition in the awake state,during anesthesia with 1.2%isoflurane,and after the end of anesthesia was 100,600 and 600 s,respectively.Imaging data were analyzed using Image J and MATLAB softwares.Results The overall trend of activity of layer 2/3 cortical neurons decreased first and then stabilized with the inhalation of 1.2%isoflurane.The cortical neural activity were gradually increased when isoflurane inhalation was stopped.The recovery rate of neural activity was different in different brain regions after isoflurane inhalation was stopped.The recovery of neural activity in the primary motor cortex was delayed obviously.During the maintenance of anesthesia,the activities of most layer 2/3 cortical neurons in the retrosplenial cortex were weakened,however,some of the neurons became more active.Conclusions The neural activity in the 2/3 layer of cortex in isoflurane anesthetized mice is inconsistent in observation region,brain region and single cell,suggesting that different neural pathways are involved in the process of anesthesia induction and recovery from anesthesia.

关 键 词：异氟醚 大脑皮质 神经元 显微观测 荧光 

分 类 号：R614[医药卫生—麻醉学]