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电化学监测胞外分泌的模拟:缓冲扩散作用下细胞-电极缝隙内的 pH 值适度变化。

Simulations of amperometric monitoring of exocytosis: moderate pH variations within the cell-electrode cleft with the buffer diffusion.

机构信息

CNRS-UMR 7010 Institut de Physique de Nice, Université Nice Côte d'Azur, Av. Joseph Vallot, 06100, Nice, France.

PASTEUR, Département de chimie, École normale supérieure, PSL University, Sorbonne Université, CNRS, 75005, Paris, France.

出版信息

Anal Bioanal Chem. 2021 Nov;413(27):6769-6776. doi: 10.1007/s00216-021-03443-z. Epub 2021 Jun 13.

DOI:10.1007/s00216-021-03443-z
PMID:34120197
Abstract

Amperometry with ultramicroelectrodes is nowadays a routine technique to investigate neurotransmitter secretion by vesicular exocytosis at the single-cell level. This electroanalytical tool allows one to understand many aspects of the vesicular release in terms of mechanisms. However, the electrochemical detection relies on the oxidation of released neurotransmitters that produce 2H and thus the possible acidification of the cell-electrode cleft. In a previous work, we considered a model involving the H diffusion or/and its reaction with buffer species. In this article, we report a more general model which takes into account the ability of buffer species to move and to be regenerated within the cell-electrode cleft. As a consequence, the pH within the cleft is still equal to its physiological value regardless of the electrochemical detection of the vesicular release for usual exocytotic cell frequencies. This confirms that amperometry at the single-cell level is a very robust technique for investigating vesicular exocytosis.

摘要

目前,微电极安培法是一种用于研究单细胞水平囊泡胞吐作用释放神经递质的常规技术。这种电分析工具可以从机制方面理解囊泡释放的许多方面。然而,电化学检测依赖于释放的神经递质的氧化,这会产生 2H,从而导致细胞-电极间隙可能酸化。在之前的工作中,我们考虑了一个涉及 H 扩散及其与缓冲物质反应的模型。在本文中,我们报告了一个更一般的模型,该模型考虑了缓冲物质在细胞-电极间隙内移动和再生的能力。因此,无论电化学检测囊泡释放的频率如何,间隙内的 pH 值仍与其生理值相等。这证实了单细胞水平的安培法是一种非常稳健的技术,可用于研究囊泡胞吐作用。

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Front Chem. 2021 Jan 11;8:591311. doi: 10.3389/fchem.2020.591311. eCollection 2020.
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Electrochemical Quantification of Neurotransmitters in Single Live Cell Vesicles Shows Exocytosis is Predominantly Partial.电化学定量分析单个活细胞囊泡中的神经递质表明胞吐作用主要是部分的。
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Utilization of HEPES for Enhancing Protein Transfection into Mammalian Cells.
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Biophys Chem. 2018 Apr;235:48-55. doi: 10.1016/j.bpc.2018.02.004. Epub 2018 Feb 8.
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The evidence for open and closed exocytosis as the primary release mechanism.作为主要释放机制的开放型和封闭型胞吐作用的证据。
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