分化早期胚胎大鼠神经干细胞电生理特征.doc

分化早期胚胎大鼠神经干细胞电生理特征 冷 洁，蒋 莉，陈恒胜 （重庆医科大学儿童医院神经内科，重庆 400014） 摘 要：目的 探索分化早期神经干细胞的电生理特征。方法 分离培养胚胎大鼠海马神经干细胞。应用膜片钳全细胞记录技术检测神经干细胞和分化早期（1～3 d）神经细胞的电压门控离子电流。结果 未分化的神经干细胞不表达内向钠离子电流，而诱导分化1 d的神经细胞即表达内向性钠离子电流，但不能诱导产生动作电位。神经干细胞和分化早期的神经细胞均表达两种类型的外向钾离子电流，并表达内向T-型钙电流。结论 钠离子电流的功能性表达是神经干细胞退出细胞周期开始分化的标志。分化早期是神经干细胞离子通道发育过程的重要阶段。 关键词：膜片钳；海马；神经干细胞；分化；离子通道 中图法分类号：R322.81;R329.2;R338.8 文献标识码：A The early electrophysiological properties of differentiated embryonic neural stem cells LENG Jie; JIANG Li; CHENG Heng-Sheng (Department of Neurology, Children’s Hospital, Chongqing Medical University, Chongqing, 400014, China) Abstract：Obiective To exam the early electrophysiological properties of cells differentiated from rat hippocampal neural stem cells. Methods Embryonic rat hippocampal neural stem cells（NSCs） were isolated and cultured. Passaged cells were plated on poly-llysine hydrochloride glass coverslips for 1 hour or differentiated for 1 to 3 days with differentiation medium, using whole cell patch-clamp technique to record the voltage-gated ion channel currents. Results No inward Na+ current was detected in any of the undifferentiated NSCs.1 d after the induction of differentiating, cells exhibited voltage-gated Na+ currents. But they were too low to generate a fast action potential. The presence of two types of outward K+ currents, delayed recitifier K+ currents and transient K+ currents and the inward T-type Ca2+ currents were also recorded in both of the NSCs and the differentiating cells. Conclusion The study demonstrated the properties of NSCs: expression two types of outward K+ currents and one type of T-type Ca2+ currents with absence of inward Na+ currents. Na+ current expression is a very early event following cell cycle exit in neurogenesis. The early stage of NSCs differentiation is critical for ion channels development. Key words:Patch-clamp techniques; hippocampus; neural stem cells; differentiation; ion channels 神经干细胞[1]，也被认为是有可能实现脑损伤后结构与功能重塑的“种子细胞”。因此对NSCs