Acid-sensing ion channels contribute to the increase in vesicular release from SH-SY5Y cells stimulated by extracellular protons.
Acid-sensing ion channels contribute to the increase in vesicular release from SH-SY5Y cells stimulated by extracellular protons.
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DOI：
10.1152/ajpcell.00067.2012
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发表时间：
2012-08
期刊：
影响因子：
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通讯作者：
Qiu-ju Xiong;Zhuang‐li Hu;Peng-Fei Wu;Lan Ni;Zhi-fang Deng;Wen-Ning Wu;Jian-Guo Chen;F. Wang
中科院分区：
文献类型：
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作者：
Qiu-ju Xiong;Zhuang‐li Hu;Peng-Fei Wu;Lan Ni;Zhi-fang Deng;Wen-Ning Wu;Jian-Guo Chen;F. Wang
酸敏感离子通道(ASIC)已被报道在神经元多巴胺途径中发挥作用，但其在神经递质释放中的确切作用仍不清楚。人神经母细胞瘤SH-SY5Y是一种能释放单胺类神经递质的多巴胺能神经元细胞系。本研究鉴定了ASICs在SH-SY5Y细胞中的表达，以进一步探讨ASICs在酸刺激的囊泡释放中的作用。我们收集的证据表明，在SH-SY5Y细胞中可以检测到ASIC。在全细胞膜片钳记录中，细胞外pH的迅速降低可诱发内向电流，该电流可被100μM阿米洛利可逆地抑制。该电流具有pH依赖性，其半最大激活(pH(0.5))为6.01±0.04。钙显影和FM1-43染料标记显示，胞外质子可增加SH-SY5Y细胞内钙离子水平和囊泡释放，这种作用可被PcTx1和阿米洛利减弱。有趣的是，N型钙通道阻滞剂抑制了酸化诱导的囊泡释放。综上所述，ASICs在SH-SY5Y细胞中有功能表达，并参与酸化刺激的囊泡释放。N型钙通道可能参与了酸诱导的囊泡释放增加。我们的结果为SH-SY5Y细胞ASICs和神经递质的释放提供了初步的研究，有助于进一步研究ASICs与多巴胺能神经元的关系。
Acid-sensing ion channels (ASICs) have been reported to play a role in the neuronal dopamine pathway, but the exact role in neurotransmitter release remains elusive. Human neuroblastoma SH-SY5Y is a dopaminergic neuronal cell line, which can release monoamine neurotransmitters. In this study, the expression of ASICs was identified in SH-SY5Y cells to further explore the role of ASICs in vesicular release stimulated by acid. We gathered evidence that ASICs could be detected in SH-SY5Y cells. In whole cell patch-clamp recording, a rapid decrease in extracellular pH evoked inward currents, which were reversibly inhibited by 100 μM amiloride. The currents were pH dependent, with a pH of half-maximal activation (pH(0.5)) of 6.01 ± 0.04. Furthermore, in calcium imaging and FM 1-43 dye labeling, it was shown that extracellular protons increased intracellular calcium levels and vesicular release in SH-SY5Y cells, which was attenuated by PcTx1 and amiloride. Interestingly, N-type calcium channel blockers inhibited the vesicular release induced by acidification. In conclusion, ASICs are functionally expressed in SH-SY5Y cells and involved in vesicular release stimulated by acidification. N-type calcium channels may be involved in the increase in vesicular release induced by acid. Our results provide a preliminary study on ASICs in SH-SY5Y cells and neurotransmitter release, which helps to further investigate the relationship between ASICs and dopaminergic neurons.