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Receptor-Mediated Field Effect Transistor Biosensor for Real-Time Monitoring of Glutamate Release from Primary Hippocampal Neurons

机译:受体介导的场效应晶体管生物传感器,用于从原发性海马神经元实时监测谷氨酸释放

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摘要

Glutamate, one of the most important central excitatory neurotransmitters, plays crucial roles in nerve signal transduction and is implicated in several neurological disorders. However, no effective means has been developed for specific detection of glutamate released from primary cultured neurons. Here we present a reduced graphene oxide (RGO)-based field effect transistor (FET) biosensor functionalized with synthesized glutamate receptor for real-time monitoring of glutamate release from primary cultured rat hippocampus neurons. Metabotropic glutamate receptors (mGluR) was specifically synthesized and then immobilized on the RGO surface by 1-pyrenebutanoic acid succinimidyl ester (PASE) linker, after which target glutamate (pI = 3.22) could specifically bind to the synthesized mGluR in the neutral buffer, causing the charge density change. After the neurons were cultured on the sensing channel with a self-made liquid reservoir, the FET biosensor could discriminate glutamate in the femtomolar range in complete cell culture medium and generate encouraging results in real-time monitoring of glutamate release from primary rat hippocampus neurons. This work is the first report of specific and direct detection of glutamate molecules released from primary culture of differentiated central neurons, which may further help understand the nature of neuronal communication. Moreover, this work paves a way for the detection of electrochemically inactive small molecules released by cells.
机译:谷氨酸是最重要的中枢兴奋性神经递质之一,在神经信号转导中起着至关重要的作用,并涉及多种神经障碍。然而,没有开发出有效的手段用于从原发性培养神经元释放的谷氨酸的特异性检测。在这里,我们介绍了用合成的谷氨酸受体官能化的石墨烯氧化物(RGO)的基础效应晶体管(FET)生物传感器,用于从原发性培养的大鼠海马神经元实时监测谷氨酸释放。特异性合成代谢谷氨酸受体(MGLUR),然后通过1-芘丁二酸琥珀酰亚胺酰亚胺(PASE)接头固定在RGO表面上,之后靶谷氨酸(PI = 3.22)可以与中性缓冲液中的合成MGLUR特异性结合,导致充电密度变化。在用自制液体贮存器上培养神经元在感测通道上培养后,FET生物传感器可以在完全细胞培养基中的毫微微罗拉范围内区分谷氨酸,并产生令人鼓舞的导致从原发大鼠海马神经元的谷氨酸释放的实时监测。这项工作是从分化的中枢神经元的原发性培养物中释放的谷氨酸分子的具体和直接检测,这可能进一步有助于了解神经元通信的性质。此外,这项工作铺设了一种方法,用于检测细胞释放的电化学无活性小分子。

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  • 来源
    《Analytical chemistry》 |2019年第13期|共8页
  • 作者单位

    Hubei Univ Chinese Med Sch Lab Med 1 Huangjia Lake West Rd Wuhan 430065 Hubei Peoples R China;

    Hubei Univ Chinese Med Sch Lab Med 1 Huangjia Lake West Rd Wuhan 430065 Hubei Peoples R China;

    Hubei Univ Chinese Med Sch Lab Med 1 Huangjia Lake West Rd Wuhan 430065 Hubei Peoples R China;

    Hubei Univ Chinese Med Clin Sch Tradit Chinese Med 1 Huangjia Lake West Rd Wuhan 430065 Hubei Peoples R China;

    Peking Univ Dept Elect Key Lab Phys &

    Chem Nanodevices 5 Yiheyuan Rd Beijing 100871 Peoples R China;

    Peking Univ Dept Elect Key Lab Phys &

    Chem Nanodevices 5 Yiheyuan Rd Beijing 100871 Peoples R China;

    Hubei Univ Chinese Med Sch Lab Med 1 Huangjia Lake West Rd Wuhan 430065 Hubei Peoples R China;

    Hubei Univ Chinese Med Sch Lab Med 1 Huangjia Lake West Rd Wuhan 430065 Hubei Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
  • 关键词

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