Real‐time Monitoring of Exocytotic Glutamate Release from Single Neuron by Amperometry at an Enzymatic Biosensor
Real‐time Monitoring of Exocytotic Glutamate Release from Single Neuron by Amperometry at an Enzymatic Biosensor
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DOI：
10.1002/elan.201700656
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发表时间：
2018-06
期刊：
影响因子：
3
通讯作者：
Quan-Fa Qiu;Fu-Li Zhang;Yun Tang;Xin-Wei Zhang;Hong Jiang;Yan-ling Liu;Weihua Huang
中科院分区：
文献类型：
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作者：
Quan-Fa Qiu;Fu-Li Zhang;Yun Tang;Xin-Wei Zhang;Hong Jiang;Yan-ling Liu;Weihua Huang
谷氨酸（Glu）是一种重要的神经递质，通过囊泡胞吐作用释放，在许多生理过程中发挥重要作用。微电极电流分析法已成为研究通过胞吐分泌的电活性生化物质的有力方法，然而，迄今为止，还没有直接监测非电活性Glu的胞吐的手段。本文中，我们通过在碳纤维微电极表面上共修饰谷氨酸氧化酶（GluOx）和Pt纳米颗粒（Pt NPs）来开发酶微生物传感器。Glu通过GluOx酶促转化为电活性产物H2O2，然后通过Pt NPs电化学检测。酶的催化活性和微米级的微电极使传感器具有高灵敏度和时空分辨率，能够真实的实时监测单个海马静脉曲张的Glu胞吐。结果表明，该生物传感器在监测和定量的神经递质谷氨酸的量子释放的能力，表明其在探索神经元通信和突触可塑性的细胞机制的巨大潜力。
Glutamate (Glu) is recognized as a vital neurotransmitter released via vesicular exocytosis and plays important roles in many physiological processes. Microelectrode amperometry has become a powerful method to study the secreted electroactive biochemicals via exocytosis, however, there was no means for direct monitoring of exocytosis of the non‐electroactive Glu so far. Herein, we developed an enzymatic micro‐biosensor by co‐modification of glutamate oxidase (GluOx) and Pt nanoparticles (Pt NPs) on the surface of a carbon fiber microelectrode. The Glu was enzymatically transformed by GluOx to an electroactive product, H2O2, which was then electrochemically detected by the Pt NPs. The prominent catalytic activity of enzyme and micrometer scale of microelectrode endow the sensor with high sensitivity and spatio‐temporal resolution, allowing real‐time monitoring Glu exocytosis from single hippocampal varicosities. The results demonstrate the capability of this biosensor in monitoring and quantification of the quantal release of neurotransmitter Glu, indicating its great potential in exploring the cellular mechanism of neuron communication and synaptic plasticity.