光纤成像技术记录小鼠眶额皮层奖赏相关神经元活性的应用  

Application of Fiber Photometry for Reward-related Neuronal Activity in the Orbital Frontal Cortex(OFC) of Mice

作　　者：刘敏[1] 孙晓红[1] 杨子成 杜庚 雷慧萌 LIU Min;SUN Xiao-hong;YANG Zi-cheng;DU Geng;LEIHui-meng(Department of eurobiology. School of Basic Medicine, Capital Medical University, Beijing, 100069, China)

机构地区：[1]首都医科大学基础医学院,北京100069

出　　处：《现代生物医学进展》2019年第8期1420-1424,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(31171051;31371108);北京市自然科学基金项目(5132007;5112008);北京市教委科技发展项目(KM201110025001)

摘　　要：目的:探讨光纤成像技术用于记录小鼠眶额皮层奖赏相关神经元活性变化的可行性。方法:应用光纤成像的方法记录自由活动小鼠在饮用糖水时,携带有钙离子荧光探针(GCaMP6m)的眶额皮层奖赏相关神经元的活性。首先,在小鼠的眶额皮层注射携带GCaMP6m的腺相关病毒,同时在相应位点植入提前做好的光纤陶瓷插芯;等待小鼠术后恢复,病毒表达2周。然后在记录前,给予小鼠36小时禁水处理并运用光纤成像记录接受糖水刺激的小鼠眶额皮层锥体神经元的反应活性。最后,记录数据读入matlab软件进行数据分析并对小鼠进行心脏灌流、取脑、脑组织冰冻切片并显微荧光成像观察记录位点是否正确,病毒是否正常表达。结果:成功记录到对小鼠施加糖水刺激时,其眶额皮层内与奖赏相关的神经元活性变化。数据分析结果用热度图和事件相关的平均线图来表示。组织学切片及成像结果证实记录位点正确,病毒正常表达。结论:光纤成像的记录方法可以监测自由活动的小鼠在饮用糖水时眶额皮层内奖赏相关神经元活性的变化。Objective: To investigate the feasibility of fiber photometry technology in recording the activity changes of reward-related neurons in the orbital frontal cortex of mice. Methods: The fiber photometry method was used creatively to record the activity of reward-related neuronal that expressed calcium fluorescent probe(GCaMP6m) in orbital frontal cortex of freely moving mice which were stimulated by drinking syrup. Firstly, adeno-associated virus carrying GCaMP6m were injected into the OFC of postanesthetic mice precisely. Then, one pre-made fiber-optic ceramic ferrule was implanted into the corresponding site carefully,accurately and suitably. The mice that undergone mentioned surgery needed to regain them health and the injected viruses needed to express GCaMP6m efficiently in two weeks. In the second place, the mice that recovered health after surgery were given water-deprivation treatment which sustained 36 hours before recorded reward-related neuronal signal. The reaction activity of frontal cortex pyramidal neurons in mice that stimulated by syrup was recorded by means of fiber photometry system dynamically. In the end,the date that reflected the activity of recorded reward-related neuro in mice’s OFC were analyzed scrupulously using matlab software. At the same time, the anaesthetic mice were subjected to heart perfusion, brain taken, frozen section of removed brain and fluorescence imaging using fluorescence microscope orderly and feelinglessly. The purpose of above complex operations were to affirm the validity of neuronal signal recording site and the level of injected viruese expression. Results: The variation of neuronal activity which associated with reward in OFC of syrup stimulated mice was recorded successfully and clearly using fiber photometry method. The results of data analysis that reflected changed neuronal activity were shown through a heat map and an event-related average plot scientifically.Meantime, the frozen section of mice brain and fluorescence imaging results all demonstrably indic

关 键 词：光纤成像 GCaMP6m 眶额皮层 奖赏 

分 类 号：R-33[医药卫生] R338