成人神经干细胞脑内移植后突触形成和电生理功能的实验研究

Synapse Formation and Electricity Physiology of Adult Human Neural Stem Cells Following Transplantation

目的研究成人神经干细胞(NSCs)脑内移植后新生神经元突触形成和电生理功能。方法从开放性脑损伤患者外露的脑组织中分离培养出成人NSCs,利用病毒基因转移的方法对成人NSCs进行标记。将rAAV-LacZ标记的成人NSCs进行裸鼠脑内移植后行免疫电镜检查,观察新生神经元突触形成情况;重组GFP逆转录病毒标记成人NSCs裸鼠脑内移植后进行脑片的细胞电生理检测。结果免疫电镜检查表明成人NSCs移植后分化的神经元与宿主细胞之间形成突触联系,膜片钳检测显示由成人NSCs分化而来的GFP阳性神经元具有自发的动作电位,全细胞膜片钳模式下可记录到Na+、K+电流。结论成人NSCs移植后可以与宿主细胞形成突触联系,其分化的神经元细胞具有电生理功能。

Objective To investigate synapse formation and electricity physiology following transplantation of adult human neural stem cells（NSCs）. Methods Adult human NSCs were isolated and propagated from exposed brain tissue of patients with open brain injury,and were labeled by rAAV-mediated LacZ gene transduction and retrovirus-mediated green fluorescent protein （GFP） transduction. The synaptic structure and electricity physiology of NSCs were investigated following transplantation 2 months later. Results Synaptic contacts were formed in grafted cells derived from adult human NSCs with host brain cells of nude mice. And those GFP positive cells from adult human NSCs processed spontaneous electricity activities,and Na^＋ , K^＋ currents were recorded by whole cell patch clamp in GFP positive cells. Conclusion The electrophysiological properties of the neurons derived from GFP-labeled adult human NSCs suggested that these new generated neurons were functional in some certain aspects.