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采用开碳纳米管定量细胞内单囊泡儿茶酚胺浓度,揭示 L-多巴对囊泡结构的影响。

Quantifying Intracellular Single Vesicular Catecholamine Concentration with Open Carbon Nanopipettes to Unveil the Effect of L-DOPA on Vesicular Structure.

机构信息

Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 41296, Gothenburg, Sweden.

出版信息

Angew Chem Int Ed Engl. 2022 Jan 3;61(1):e202113406. doi: 10.1002/anie.202113406. Epub 2021 Nov 23.

Abstract

Understanding the regulatory mechanisms of exocytosis is essential for uncovering the pathologies of neuronal disorders and developing related pharmaceuticals. In this work intracellular vesicle impact electrochemical cytometry (IVIEC) measurements with different-sized (50-500 nm radius) open carbon nanopipettes (CNPs) were performed to quantify the vesicular content and release kinetics of specific vesicle populations grouped by orifice sizes. Intracellular vesicles with radius below 100 nm were captured and narrowed between 50 and 100 nm. On the basis of this, single vesicular catecholamine concentrations in the intracellular environment were quantified as 0.23-1.1 M. Our results with L-3,4-dihydroxyphenylalanine (L-DOPA)-exposure indicate that L-DOPA regulates exocytosis by increasing the dense core size and vesicular content while catecholamine concentrations did not show obvious alterations. These were all achieved simultaneously and relatively noninvasively with open CNPs.

摘要

了解胞吐作用的调控机制对于揭示神经元疾病的发病机制和开发相关药物至关重要。在这项工作中,使用不同尺寸(半径 50-500nm)的开碳纳米管(CNP)进行细胞内囊泡冲击电化学细胞术(IVIEC)测量,以定量分析按孔径大小分组的特定囊泡群体的囊泡内容物和释放动力学。半径小于 100nm 的细胞内囊泡被捕获并缩小至 50-100nm。在此基础上,定量分析了细胞内环境中单囊泡儿茶酚胺的浓度为 0.23-1.1M。我们用 L-3,4-二羟苯丙氨酸(L-DOPA)暴露的结果表明,L-DOPA 通过增加致密核心的大小和囊泡内容物来调节胞吐作用,而儿茶酚胺浓度没有明显改变。所有这些都是用开碳纳米管同时且相对无创地实现的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b74/9299131/679f14e7a1bb/ANIE-61-0-g003.jpg

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