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目的研究离体培养的海马神经元LKB1表达下调对于神经元微小兴奋性突触后电流(mEP-SC)的影响。方法选用17d的胚胎大鼠培养海马神经元,分别用电穿孔的方法转染CAG-RE质粒和LKB1RNAi质粒,培养10~12d后进行电生理记录,选用全细胞膜片钳方式及自由记录模式,细胞外液加TTX阻断动作电位,加Bicucullin抑制GABA电流,记录神经元的mEPSC,比较2组神经元的mEPSC频率和幅度的差别。结果转染CAG-RE的神经元mEPSC幅度平均为25.6pA,频率平均为(5.21±0.25)Hz,是基线的99.8%;转染LKB1RNAi的神经元mEPSC幅度平均为35.1pA,频率平均为(5.79±0.27)Hz,是基线的127.1%;比较2组间频率、幅度变化,差别有显著性意义(P<0.05)。结论LKB1基因表达下调增强了培养海马神经元突触传递的效率。
Objective To investigate the effect of down-regulation of LKB1 in cultured hippocampal neurons on mEP-SC of neurons. Methods The hippocampal neurons were cultured in 17-day-old embryos of rats. The plasmids of CAG-RE plasmid and LKB1 RNAi were transfected by electroporation. After cultured for 10-12 days, electrophysiological recordings were performed. Whole-cell patch clamp and free- External fluid plus TTX blocking action potential, plus Bicucullin inhibition of GABA current, recording mEPSC neurons, mEPSC neurons in two groups compared the frequency and amplitude differences. Results The average mEPSC amplitude of neurons transfected with CAG-RE was 25.6pA and the average frequency was (5.21 ± 0.25) Hz, which was 99.8% of that of baseline. The mean mEPSC amplitude of neurons transfected with LKB1RNAi was 35.1pA and the average frequency was 5.79 ± 0.27) Hz, which was 127.1% of the baseline. Comparing the frequency and amplitude of the two groups showed significant difference (P <0.05). Conclusion Down-regulation of LKB1 gene enhances the efficiency of synaptic transmission in cultured hippocampal neurons.