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目的探讨酸敏感离子通道(ASICs)在海马神经元树突发育中的作用。方法在体外培养第5天的原代海马神经元中转染定位于膜上的绿色荧光蛋白(F-GFP),随后在神经元培养液中加入ASICs拮抗剂Amiloride和ASIC1a选择性拮抗剂Psalmotoxin 1(PcTX1)抑制ASICs的功能,观察体外培养8d和14d这两个时间点海马神经元的树突生长、分支复杂程度。结果 Amiloride(10-5mol/L)和PcTX1(1∶20 000稀释)处理3d对海马神经元树突分支总长度、树突分支总数均无显著影响,表明在海马神经元发育早期短时间抑制ASICs功能不影响树突发育。Amiloride(10-5mol/L)处理9d可以显著降低树突分支总长度和树突分支总数;PcTX1(1∶20 000稀释)处理9d也可以显著降低树突分支总长度,但对树突分支总数无显著影响。实验结果表明,长时间抑制ASICs功能会影响树突发育。结论 ASICs参与调节树突的发育。
Objective To investigate the role of acid sensitive ion channels (ASICs) in the dendrites of hippocampal neurons. Methods The green fluorescent protein (F-GFP) located on the membrane was transfected into primary cultured hippocampal neurons on day 5, and then the ASICs antagonist Amiloride and ASIC1a selective antagonist Psalmotoxin 1 (PcTX1) inhibits the function of ASICs. The dendritic growth and branch complexity of hippocampal neurons cultured at 8 and 14 days in vitro were observed. Results Amidoride (10-5mol / L) and PcTX1 (1: 20000 dilution) did not affect the total length of dendritic branches and total number of dendritic branches in hippocampal neurons, indicating that inhibition of ASICs Function does not affect dendritic development. Amiloride (10-5mol / L) for 9d significantly reduced the total length of dendritic branches and the total number of dendritic branches; PcTX1 (1: 20000 dilution) for 9d also significantly reduced the total length of dendritic branches, but the total number of dendritic branches No significant effect. The experimental results show that long-term inhibition of ASICs function will affect dendritic development. Conclusion ASICs are involved in the regulation of dendritic development.