光基因技术选择性调控星形胶质细胞内钙离子水平的研究  

Effect of Selective Optogenetic Regulation on Astrocyte Intracellular Calcium

作　　者：魏文洁 苏醒 谢敏杰[1] 王伟[1] 骆翔 喻志源[1] WEI Wen-jie;SU Xing;XIE Min-jie;WANG Wei;LUO Xiang;YU Zhi-yuan(Department of Neurology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Department of Oncology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030 [2]华中科技大学同济医学院附属同济医院肿瘤科,武汉430030

出　　处：《神经损伤与功能重建》2020年第4期187-189,197,共4页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金面上项目(No.81771341);国家自然科学基金青年项目(No.81600990);湖北省自然科学基金(No.2017CFB705)。

摘　　要：目的:研究光敏感通道蛋白(ChR2)选择性调控星形胶质细胞,对其细胞内钙离子水平的影响。方法:离体培养SD大鼠星形胶质细胞细胞,采用慢病毒载体Lenti-GFAP-ChR2-YFP感染星形胶质细胞后,钙离子荧光探针Rhod-2 AM作为钙离子活动的指示,以470 nm波长的蓝光刺激星形胶质细胞,实时在共聚焦荧光下检测星形胶质细胞钙离子活动。结果:成功将光敏感通道蛋白(ChR2)表达在星形胶质细胞;激活光敏感通道ChR2后,引起星形胶质细胞内钙信号活动,钙离子水平增加。结论:以光敏感通道ChR2为核心的光基因技术可运用于调控星形胶质细胞钙离子活动。Objective:To study the effect of selective optogenetic regulation by Channelrhodopsin2(ChR2)on intracellular calcium levels in astrocytes.Methods:Astrocytes were cultured from SD neonate rats,and a lentiviral vector,Lenti-GFAP-ChR2-YFP,was used to transduce cultured astrocytes.Astrocytes were stimulated using blue light with a wavelength of 470 nm,and intracellular calcium signals,as indicated by the fluorescent Ca2+indicator Rhod-2 AM,were recorded in real time by confocal fluorescence microscopy.Results:ChR2 was successfully expressed in astrocytes.Photostimulation of ChR2 in astrocytes led to the activation of intracellular calcium signaling and an increase in calcium levels.Conclusion:The use of ChR2 as an essential element of optogenetics could be a powerful tool in the regulation of calcium in astrocytes.

关 键 词：光基因技术 光敏感通道2 星形胶质细胞 钙离子 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学]