不同脑区注射慢病毒对大鼠齿状回神经元的影响

Effects of lentiviral delivery in different brain regions on transfection of rat dentate gyrus neurons

目的探讨在不同脑区（齿状回、海马CA1和内嗅皮质）注射慢病毒对大鼠海马齿状回部位神经元的转染效果。方法健康雄性Sprague Dawley大鼠随机分为齿状回（dentate gyms，DG）组、海马CA1（Cornu Ammonis 1）组、内嗅皮质（medial entorhinal cortex，EC）组和DG—EC组，每组5只。所有实验大鼠均在脑立体定位仪上定位注射慢病毒Lenti-pSyn-GFP（5μl）。慢病毒注射2周后，4％多聚甲醛灌注、固定、取脑，30μm冰冻切片，行Nissl荧光染色，观察大鼠DG部位绿色荧光蛋白（GFP）的表达情况。结果DG、CA1、EC、DG—EC组大鼠齿状回部位绿色荧光蛋白（GFP）阳性细胞数分别为（18．0±1．0）个／视野，（17．0±0．6）个／视野，（17．3±0．6）个／视野，（18．3±0．6）个／视野，各组间差异无统计学意义（均P〉0．05）。结论在大鼠CA1或内嗅皮质部位注射慢病毒后均能够转染DG部位的神经元，代替直接在DG部位进行注射转染DG区神经元，从而减少对DG部位的损伤。

Objective To explore the effect of lentiviral delivery green fluorescent protein in different brain regions on the transfection of dentate gyms neurons. Methods Lenti-pSyn-EGFP was stereotaxic injected into dentate gyms （ DG）, Cornu Ammonis 1 （ CA1 ） of hippocampus and medial entorhinal cortex （EC） of rats. All animals were perfused with 4% paraformaldehyde. Their brains were cut into 30 μm sections which were performed with Nissl fluorescent staining. Results The resuts showed that the GFP positive cells in DG area of DG,CA1 ,EC,DG-EC groups were （ 18.0±1.0）, （ 17.0±0.6）, （ 17.3±0.6）, （18.3±0.6） respectively,and there was no statistical difference among each group（P〉0.05）. Conclusion Lentivirus delivery in EC and CA1 both can transfect DG neurons. This is a useful method to transfect dentate gyms neurons instead of injecting lentivirus into DG directly and avoid the lesion of DG.