一种特异性靶向神经节神经元的基因递送技术

A gene delivery technique to specifically target ganglion neuron

目的开发一种无创条件下于外周神经系统表达腺相关病毒(adeno-associated virus,AAV)以特异性操纵神经节神经元的方法。方法出生0天的小鼠腹腔注射10μL AAV2/9-EGFP,病毒滴度为(2~5)×10^(12)vg/mL,待小鼠长至2月龄时灌注取各组织、器官切片,免疫荧光染色,观察增强绿色荧光蛋白(enhanced green fluorescent protein,EGFP)的亮度及范围,判断病毒感染情况。结果69.11%±1.42%的背根神经节(dorsal root ganglion,DRG)神经元,7.08%±1.16%的三叉神经节(trigeminal ganglion,TG)神经元,32.46%±1.81%的迷走神经节(nodose ganglion,NG)神经元和4.76%±0.29%的交感神经节(sympathetic ganglion,SG)神经元被感染。脑、脊髓、内脏器官中的神经元被感染效率极低。结论本方法可特异性感染神经节神经元,但对神经胶质细胞、中枢神经系统神经元或外周器官感染效率极低,可用于特异性操纵神经节神经元。

Objective To develop a noninvasive method for expressing adeno-associated virus(AAV)in the peripheral nervous system to specifically manipulate ganglionic neurons.Methods P0 mice were intraperitoneally injected 1012μL AAV2/9-EGFP with a viral titer of(2-5)×10^(12) vg/mL.When the mice were 2 months old,the tissues and organs were perfused and cut into slices.Immunofluorescence staining was performed on sections to observe the brightness and range of enhanced green fluorescent protein(EGFP)and determine the virus infection.Results 69.11%±1.42%dorsal root ganglion(DRG)neurons,7.08%±1.16%trigeminal ganglion(TG)neurons,32.46%±1.81%nodose ganglion(NG)neurons,and 4.76%±0.29%sympathetic ganglion(SG)neurons were infected,while neurons in brain,spinal cord and internal organs were rarely infected.Conclusion This method can specifically infect ganglionic neurons,but has extremely low efficiency in infecting glial cells,central nervous system neurons,or peripheral organs.It can be used to specifically manipulate ganglionic neurons.