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机构地区:[1]温州医学院眼视光学院,卫生部视觉科学重点实验室,浙江温州325027
出 处:《解剖学报》2012年第2期184-188,共5页Acta Anatomica Sinica
基 金:浙江省自然科学基金(Y2100855);温州市科技局科技计划项目(Y20100126);温州医学院附属眼视光医院科研启动基金(KYQD090701)
摘 要: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-sensing ion channels (ASICs) in dendritic development in hippocampal neurons. Methods F-GFP which targets at the membrane was transfected into primary cultured hippocampal neurons at the 5th days in vitro (DIV 5). Then ASICs antagonist amiloride and ASICla selective antagonist psalmotoxin 1 (PcTX1) were applied to the neuronal medium to inhibit the function of ASICs. Dendritic growth and arborization of cultured hippocampal neurons were observed in DIV8 and DIV14. Results Treatments with ASICs antagonist amiloride ( 10 -Stool/L) and ASICla selective antagonist PcTX1 ( 1:20 000 dilution) for 3 days had no significant effect on the total dendritic branch length and total dendritic branch number of hippocampal neurons, which suggested that dendritic development was not affected by the inhibition of ASICs for a short time during early developmental period of hippocampal neurons. Chronic treatment with amiloride (10-5mol/L) for 9 days significantly reduced the total dendritic branch length and total dendritic branch number, while chronic treatment with ASICla selective antagonist PeTX1 (1:20 000 dilution) for 9 days also significantly reduced the total dendritic branch length, but had no significant effect on the total dendritic branch number, which suggested that dendritic development was impaired if ASICs are inhibited for a long time. Conclusion ASICs play an essential role in dendritic development.
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