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脑内转染含胶质纤维酸性蛋白启动子的慢病毒对小鼠脑电图的影响
许晓惠1,刘娟1,2,邹静1,3,王国祥2,王云2,李胜天1*
0
(1. 上海交通大学Bio-X研究院, 上海 200040;
2. 复旦大学脑科学研究院, 上海 200032;
3. 上海交通大学医学院附属仁济医院神经内科, 上海 200001
*通信作者)
摘要:
目的 本研究通过检测胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)启动子(pGFAP)慢病毒海马注射对小鼠脑电图活动的影响,探讨其在癫(疒间) 研究中的可行性。 方法 采用免疫荧光检测方法检测pGFAP慢病毒在培养海马神经元体系以及小鼠海马内对星形胶质细胞感染的特异性。将小鼠分成两组,脑内注射人工脑脊液组与脑内注射pGFAP慢病毒组,检测脑内注射前后两组小鼠体质量的变化;使用Spike 2软件分析脑内注射3~4周后两组小鼠脑电图的功率谱以及高频振荡的变化。 结果 (1)pGFAP慢病毒在培养海马神经元体系中特异性感染非神经元细胞;(2)小鼠海马齿状回(dentate gyrus,DG)区注射pGFAP慢病毒特异性感染GFAP阳性细胞;(3)与人工脑脊髓液注射组相比,小鼠体质量在病毒注射3~4周后无明显差异;(4)海马DG区pGFAP病毒注射未明显改变小鼠脑电图功率谱,高频振荡的数目和平均持续时间均无明显改变。 结论 应用pGFAP慢病毒脑内注射方法研究星形胶质细胞功能在癫(疒间) 中的作用是可行的。
关键词:  慢病毒属  神经胶质原纤维酸性蛋白  启动子  癫(疒间)  脑电描记术
DOI:10.3724/SP.J.1008.2015.1173
投稿时间:2015-03-04修订日期:2015-06-15
基金项目:上海市自然科学基金(13DJ1400303).
Effect of brain transfection of glial fibrillary acidic protein promoter-containing lentivirus on electroencephalogram activity in mice
XU Xiao-hui1,LIU Juan1,2,ZOU Jing1,3,WANG Guo-xiang2,WANG Yun2,LI Sheng-tian1*
(1. Bio-X Institute, Shanghai Jiaotong University, Shanghai 200240, China;
2. Institute of Brain Science, Fudan University, Shanghai 200032, China;
3. Department of Neurology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200001, China
*Corresponding author.)
Abstract:
Objective To analyze the effects of injecting glial fibrillary acidic protein promoter (pGFAP)-containing lentivirus into mouse hippocampus on electroencephalogram(EEG)activity, and to explore the feasibility of applying the lentivirus in epilepsy study. Methods Astrocytes-specific infection of pGFAP lentivirus was identified in both cultured hippocampal cells and hippocampal slices from mice by immunofluorescence techniques. The mice were divided into two groups: artificial cerebrospinal fluid(ACSF)injection group and pGFAP lentivirus injection group. Changes in mice body weight were analyzed before and after brain injection. Changes of power spectrum and high-frequency oscillations(HFOs)of EEG were analyzed by Spike 2 software 3 to 4 weeks after pGFAP lentivirus injection. Results It was found that:(1)non-neuron cells were selectively infected by pGFAP lentivirus in cultured hippocampal cells;(2)pGFAP lentivirus specifically infected GFAP+cells in dentate gyrus(DG)area of mouse hippocampus;(3)compared with ACSF injection group, mouse body weight had no significant changes 3 to 4 weeks after the virus injection in pGFAP lentivirus injection group; and(4)pGFAP lentivirus injected into DG area of hippocampus had no significant effect on power spectrum, or the number and mean duration of HFOs of EEG. Conclusion It is a feasible strategy to study the function of astrocytes in epilepsy by brain injection of pGFAP lentivirus.
Key words:  lentivirus  glial fibrillary acidic protein  promoter  epilepsy  electroencephalography