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目的:研究新鲜分离的新生大鼠皮层神经元胞内钙离子浓度([Ca~(2+)]_i)发生振荡的机制。方法:采用酶解结合机械分离法从6—7日龄大鼠分离皮层神经元,用M40钙离子测量系统(PTI)测量细胞内钙离子浓度的变化。用Fura-2作为钙离子指示剂。结果:在观察到的82个神经元细胞中,47个产生了自发钙振荡。自发钙振荡依赖于胞外钙离子浓度。去除外钙后自发钙振荡立即停止。四乙铵1mmol/L引起钙振荡振幅增大,频率变快。CsCl 1mmol/L主要引起频率增加。BaCl_2 1mmol/L可使振幅、频率增高,并有明显的高台样基线增加。结论:皮层神经元在无突触联系的情况下具有产生自发[Ca~(2+)]_i振荡的特性,K~+通道在决定钙振荡的幅值和频率方面起重要作用。
AIM: To investigate the mechanism of intracellular Ca2 + concentration ([Ca2 +] i) oscillations in freshly isolated neonatal rat cortical neurons. Methods: Cortical neurons were isolated from 6-7 day old rats by enzymolysis combined with mechanical separation. The changes of intracellular calcium concentration were measured by M40 calcium ion measurement system (PTI). Fura-2 as a calcium ion indicator. Results: Of the 82 neuronal cells observed, 47 produced spontaneous calcium oscillations. Spontaneous calcium oscillations depend on the extracellular calcium concentration. Remove the calcium after spontaneous calcium oscillation immediately stop. Tetraethylammonium 1mmol / L caused calcium oscillation amplitude increases, the frequency becomes faster. CsCl 1mmol / L mainly caused the increase of frequency. BaCl_2 1mmol / L can make the amplitude and frequency increased, and there was a clear increase in baseline height. CONCLUSION: Cortical neurons have spontaneous [Ca2 +] _i oscillations in the absence of synaptic connections, and K ~ + channels play an important role in determining the magnitude and frequency of calcium oscillations.