目的:研究发育过程中大鼠视皮层2/3层锥体神经元的白发兴奋性突触后电流(sEPSCs)变化,以及在生后早期自发性突触活动情况,探讨视觉经验在视皮层发育过程中对神经元突触的修饰作用。方法:应用红外微分干涉相差显微镜(m—DIC)结合电荷耦合式摄像机(CCD—Camera)可视法膜片钳全细胞记录生后2—7、8~14、15-21、22-28d各组sEPSCs变化。同时于电极内液中加入虫荧光黄,对所记录细胞进行染色观察形态学改变。结景:视皮层神经元sEPSCs幅值随发育逐渐升高(单因素方差分析P〈0.01)。视皮层神经元sEPSCs频率随发育逐渐提高(单因素方差分析P〈0.01)。但2~7d组与8—14d组差异无统计学意义(两独立样本t检验P〉0.05)。视皮层2,3层神经元胞体及突起以及生物电学特性随发育逐渐成熟。结论:大鼠视皮层锥体神经元突触后膜AMPA受体功能表达随发育逐渐成熟,生后早期突触并非完全处于静息状态,具有一定的早期白发突触功能活动。
Objective: To investigate the modificational function of visual experience onto synapses by recording developmental changes of spontaneous EPSCs in layer 2/3 pyramidal neurons of rat visual cortex, and observe the spontaneous synaptic activities during the earlier postnatal period. Methods: By combining infrared differential interference contrast (IR-DIC) technique and CCD-Camera system with visual patch clamp whole-cell recording technique, spontaneous EPSCs of P2-7,P8-14,P15-21,P22-28 groups were oberved and analyzed,simultaneously the neurons were stained by adding luciferyellow to the pipette solution. Results: The amplitude of sEPSCs was enhanced when aging (one way ANOVA, P〈0.01).The sEPSCs frequency increased with age (one way ANOVA, P〈0.01).There was no significant difference between P2-7 group and P8-14 group (two independent sample t-test, P〉0.05). The electrophysiology and dendrite of neuron became mature over development. Conclusion: Although most synapses are regarded as silent synapses at early postnatal days,which show exclusively NMDA receptor-mediated function,the results imply there are early AMPA receptor-mediated functional activities in layer 2/3 pyramidal neurons,synapses are not completely silent, they do have early spontaneous activities.