成年新生神经元NMDA受体NR2B亚型基因条件性敲除模型建立  

Establishment of Conditional NR2B Gene Knock-out Model in New-born Granule Cells in Adult Mice

作　　者：李志军[1] 何丽琳[2] 易陈菊[1] 唐娜[1] 

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]天门市第一人民医院肿瘤科,湖北天门431700

出　　处：《神经损伤与功能重建》2013年第1期1-6,共6页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金青年基金(No.81000537)

摘　　要：目的:建立成年神经发生中新生颗粒细胞N-甲基-D-天冬氨酸(NMDA)受体NR2B亚型基因条件性敲除模型。方法:运用逆转录病毒基因标记技术,结合Cre-1oxp重组酶系统,在ROSA26-LacZ基因报告小鼠体内研究逆转录病毒RSV-pGFP:T2aCre的Cre酶重组的发生以及其转化效率。在NR2Bfl/fl转基因小鼠中建立成年新生颗粒细胞NR2B基因敲除模型。结果:在逆转录病毒注射后14 d,观察到LacZ标记的神经元及β-gal和Cre免疫荧光双标阳性细胞,在海马齿状回颗粒下层、颗粒层以及hilus区表达;同时在嗅球颗粒层也可见LacZ表达。28 dCre转化效率高达98.3%。在NR2Bfl/fl转基因小鼠,14 d时分别在嗅球和齿状回的颗粒细胞层,观察到GFP和Cre同时标记的阳性新生颗粒细胞。结论:成年神经发生中起源于海马齿状回颗粒下层和侧脑室旁脑室管膜下区的新生颗粒细胞NR2B基因敲除模型建立。Objective： To establish a conditional NR2B gene knock-out model in the progenitors of granule cells generated from the subventricular zone（SVZ） and subgranular zone（SGZ） in adult neuro- genesis. Methods： Using retroviral genetic labeling approaches and Cre-loxp recombinant system, the expression of Cre gene and the recombinant efficiency have been studied in the ROSA26-LacZ gene reporter mice. Using retroviral genetic method to establish a conditional NR2B gene knock-out model in new born granule cells in adult neurogenesis in NR2Bfl/fl mice. Results： At day 14 after virus in- jection, Cre expressed in the new born granule cells in the granule cell layer and subgranule zone in the dentate gyms in hippocampus, as well as in the granule cell layer in the olfactory bulb, revealed by LacZ staining and immunofluorescence staining with GFP and ？-gal double labeling. These results demonstrated the successful recombinant of Cre enzyme in the adult born granule cells, Further result suggested that the efficiency of the Cre recombinant was around 98.3%. As we expected, in NR2Bfl/fl transgenic mice, the Cre recombinant also succeeded with the expression of GFP and Cre double la- beled adult-born cells both in olfactory bulb and hippocampus. Conclusion： The successful establish- ment of the conditional NR2B gene knock-out model in new-born granule cells in adult neurogenesis.

关 键 词：成年神经发生 NMDA受体NR2B亚型 Cre-loxp重组酶系统 条件性基因敲除 

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]