摘要
目的介绍建立体外颞叶癫痫神经元模型的方法,并且通过原子力显微镜(atomic force microscope,AFM)初步研究其表面的显微结构。方法将培养14d的神经元放入"无镁"细胞外液处理3h后,再重新放回含镁的正常细胞外液培养,利用膜片钳记录神经元的自发性电活动。将"无镁"细胞外液处理3h的神经元定为实验组,将未经"无镁"外液处理的神经元定为对照组。AFM分别在80μm×80μm,2μm×2μm和500nm×500nm扫描范围,对两组神经元细胞膜表面进行扫描,同时测量两组神经元表面相关结构的直径和深度。结果膜片钳结果表明神经元在正常细胞外液中未记录到阵发性去极化飘移(paroxysmal depolarization shift,PDS);"无镁"细胞外液处理3h和恢复正常外液14d时,神经元记录到PDS。在AFM扫描范围为80μm×80μm时,两组神经元细胞膜表面光滑;在2μm×2μm时,两组神经元细胞膜表面出现一些小凹;在500nm×500nm时,实验组神经元表面小凹的直径和深度分别为114.86±9.33nm和5.71±0.69nm,对照组为116.4±9.13nm和5.69±0.71nm,两组相比无显著性差异(P>0.05)。结论"无镁"外液颞叶癫痫神经元模型稳定、可靠;"无镁"外液处理神经元3h,神经元细胞膜显微结构未发生改变。
Objective Introducing a method to establishing the model of epileptic neuron of temperal lobe epilepsy. Methods Model of ＂epileptic neuron＂ was established by 3h magnesium -free media treatment. Patch clamp was applied to record the electrophysiology of epileptic neuron. AFM was used to observe and measure microstructure of the membrane of epileptic neuron. Results The neuron in normal medium, paroxysmal depolarization shift （PDS） was not recorded. After the 3h magnesium free media treatment, the neuron displayed PDS which had been continued after neuron returned to normal medium culture on day 14. There was not difference on membrane of neurons between epileptic neuron and normal neuron under scanning size 80μm× 80μm and 2μm× 2μm. Under scanning size 500nm× 500nm, the radium and depth of pits in treatment group was 114. 86 ±9. 33nm and 5. 71±0. 69nm respectively, and in control group was 116.4±9. 13nm and 5. 69±0. 71nm. There was not significant difference between two groups. Conclusion The model of epileptic neuron by by 3h magnesium-free media treatment was stable and reliable. There was no change in membrane of neurons suffering from 3h magnesium free media treatment.
出处
《立体定向和功能性神经外科杂志》
2008年第1期12-16,共5页
Chinese Journal of Stereotactic and Functional Neurosurgery
基金
哈医大一院院基金资助项目(编号:2007030)
关键词
原子力显微镜
癫痫神经元
膜片钳
Atomic force microscope
Epileptic neuron
Patch clamp
