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目的探讨神经干细胞在癫癎微环境下能否发育为“癫癎神经元”。方法“无镁”外液处理神经元建立“癫癎神经元”模型。将绿色荧光蛋白标记的神经干细胞分别与正常海马神经元、“癫癎神经元”共培养,应用膜片钳记录干细胞的突触后电位,利用免疫荧光检测干细胞突触素抗体染色情况。将神经干细胞分化神经元放入“无镁”外液,应用膜片钳记录其突触后电位。结果在“无镁”细胞外液处理3h 后恢复正常细胞外液培养14d,神经元仍存在自发的“癫癎样放电”。干细胞与“癫癎神经元”共培养时,免疫荧光结果示80%干细胞突触素染色阳性,膜片钳记录到干细胞12次/5min 兴奋性突触后电位。在“无镁”外液中,60%干细胞分化神经元出现14次/5min 时程约10s的兴奋性突触后电位,未记录到“癫癎样放电”。结论干细胞能够与周围神经元形成功能性突触;干细胞在癫癎微环境下转变成“癫癎神经元”的可能性极小。
Objective To investigate whether neural stem cells can develop into “epileptic neurons” under epileptic microenvironment. Methods “Magnesium-free” external fluid treatment of neurons to establish “epileptic neuron” model. Green fluorescent protein-labeled neural stem cells were co-cultured with normal hippocampal neurons and epileptic neurons. Patch clamp was used to record the postsynaptic potential of stem cells. The syncytium antibody staining of stem cells was detected by immunofluorescence. Divide neural stem cells into neurons in “magnesium-free ” external fluid, the use of patch-clamp recording of its postsynaptic potential. The results in the “magnesium-free” extracellular solution after 3h return to normal extracellular fluid culture 14d, neurons still exist spontaneous “epileptiform discharge”. When stem cells were co-cultured with “epileptic neuron ”, immunofluorescence results showed that 80% of the stem cells were positive for synaptophysin staining and 12 times for the stem cells / excitatory postsynaptic potential for 5 minutes. In the “magnesium-free” extracorporeal fluid, 60% of the stem cells differentiated neurons appeared 14 times / 5min duration of about 10s excitatory postsynaptic potential, did not record the “epileptiform discharge”. Conclusion Stem cells can form functional synapses with peripheral neurons. Stem cells are less likely to be converted into “epileptic neurons” under epileptic microenvironment.