慢病毒介导的Lhx8基因过表达在海马胆碱能神经元发生中所起作用的初步研究  

A preliminary study of lentivirus-mediated Lhx8 overexpression in hippocampal cholinergic neurogenesis

作　　者：李浩明[1] 秦建兵[1] 金国华[1] 施金洪[1] 邹琳清[1] 张新化[1] 韩笑[1] 丁会[1] 成翔[1] 

机构地区：[1]南通大学医学院人体解剖学系,南通226001

出　　处：《南通大学学报（医学版）》2014年第5期349-351,F0002,共4页Journal of Nantong University(Medical sciences)

基　　金：国家自然科学基金资助项目(31271138);江苏省自然科学基金资助项目(Bk2012659);江苏省高校自然科学研究项目(13KJB310011)

摘　　要：目的:明确Lhx8过表达慢病毒在海马齿状回胆碱能神经元发生中的作用。方法:构建Lhx8过表达重组质粒、进行慢病毒包装,通过颅脑立体定位和微量注射的方法,将所得慢病毒Lenti6.3-Lhx8注入SD大鼠海马齿状回中;于注射后第1、3、7、10、14天在激光共聚焦显微镜下直接观察增强绿色荧光蛋白(enhanced green fluorescent protein,EGFP)的荧光表达;检测术后第7天时EGFP/Lhx8/ChAT三标阳性细胞的数目。结果:慢病毒注射后第3天,海马齿状回中EGFP绿色荧光信号最强;此后EGFP表达下降,注射后第14天,几乎检测不到EGFP阳性细胞;术后第7天,注射Lenti6.3-Lhx8的慢病毒侧海马齿状回门区内能够检测到EGFP/Lhx8/ChAT三标阳性细胞,与阴性对照侧比较明显增多。结论:海马齿状回内注射Lhx8过表达慢病毒能够促进胆碱能神经元的发生。Objective： To understand the function of Lhx8 lentivirus-induced cholinergic neurogenesis in hippocampal den- tate gyrus. Methods： The Lhx8 overexpressed recombinant plasmid was constructed and the lentivirus Lenti6.3-Lhx8 was ac- quired, then Lenti6.3-Lhx8 was injected into SD rats＇ hippocampal dentate gyrus using brain stereotaxic and microinjection technique. The expression of enhanced green fluorescent protein（EGFP） was investigated by using confocal laser microscope at the 1, 3, 7, 10, 14 day after lentivirus injection, and the EGFP/LhxS/ChAT positive cells were detected on the 7 day after lentivirus injection. Results： The EGFP signal reached the peak level on the 3 day after lentivirus injection in hippocampal dentate gyrus; then the EGFP signal decreased, and no obviously EGFP positive cell could be found on the 14 day after in- jection; on the 7 day after Lenti6.3-Lhx8 injection, the EGFP/Lhx8/ChAT positive cells were detected in hilus of hippocam- pal dentate gyrus, and the cells number was more than that in the negative control side. Conclusion： The injection of Lhx8 lentivirus promotes the cholinergic neurogenesis in hippocampal dentate gyrus.

关 键 词：Lhx8 海马 慢病毒 过表达 神经元 

分 类 号：R338.26[医药卫生—人体生理学]