Neurotransmitter Readily Escapes Detection at the Opposing Microelectrode Surface in Typical Amperometric Measurements of Exocytosis at Single Cells.
Neurotransmitter Readily Escapes Detection at the Opposing Microelectrode Surface in Typical Amperometric Measurements of Exocytosis at Single Cells.
复制标题
在单细胞胞吐作用的典型安培测量中，神经递质容易在相对的微电极表面逃避检测。
DOI：
10.1021/acs.analchem.2c00060
复制
发表时间：
2022-07-12
影响因子：
7.4
通讯作者：
Sombers, Leslie A.
中科院分区：
文献类型：
--
作者：
McCarty, Gregory S.;Dunaway, Lars E.;Denison, J. Dylan;Sombers, Leslie A.
几十年来，碳纤维微电极已被用于在各种细胞类型的神经递质释放的安培测量，提供了大量的有价值的信息，涉及致密核心囊泡融合的机制。可以在相对的微电极表面检测释放的电活性分子，从而允许精确定量以及关于神经递质释放阶段的详细动力学信息。然而，目前还不清楚有多少释放到人工突触中的儿茶酚胺逃脱了检测。这项工作研究了两个独立的机制，其中释放的神经递质去检测在一个典型的安培测量。首先，扩散损失进行评估，通过监测胞吐在单个牛嗜铬细胞使用碳纤维微电极制造的凹（腔）的几何形状。这就形成了一个微量取样瓶，可在检测前最大限度地减少分析物的扩散损失。与常规配置相比，当使用凹腔传感器时，每个胞吐释放事件检测到更多的分子。此外，药理学抑制去甲肾上腺素转运蛋白（NET），这是用来从细胞外空间中去除儿茶酚胺，增加了个人安培事件的大小和时间过程。总的来说，这项研究的特点是不同的物理和生物机制，释放的神经递质逃脱检测在相对的微电极表面，同时也揭示了一个重要的作用，NET在“突触前”调制的神经递质释放。
For decades, carbon-fiber microelectrodes have been used in amperometric measurements of neurotransmitter release at a wide variety of cell types, providing a tremendous amount of valuable information on the mechanisms involved in dense-core vesicle fusion. The electroactive molecules that are released can be detected at the opposing microelectrode surface, allowing for precise quantification as well as detailed kinetic information on the stages of neurotransmitter release. However, it remains unclear how much of the catecholamine that is released into the artificial synapse escapes detection. This work examines two separate mechanisms by which released neurotransmitter goes undetected in a typical amperometric measurement. First, diffusional loss is assessed by monitoring exocytosis at single bovine chromaffin cells using carbon-fiber microelectrodes fabricated in a recessed (cavity) geometry. This creates a microsampling vial that minimizes diffusional loss of analyte prior to detection. More molecules were detected per exocytotic release event when using a recessed cavity sensor as compared to the conventional configuration. In addition, pharmacological inhibition of the norepinephrine transporter (NET), which serves to remove catecholamine from the extracellular space, increased both the size and the time course of individual amperometric events. Overall, this study characterizes distinct physical and biological mechanisms by which released neurotransmitter escapes detection at the opposing microelectrode surface, while also revealing an important role for the NET in “presynaptic” modulation of neurotransmitter release.
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DOI：
10.1083/jcb.57.2.499
发表时间：
1973-05
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ceccarelli B;Hurlbut WP;Mauro A
通讯作者：
Mauro A
影响因子：
8.4
作者：
Aref M;Ranjbari E;Romiani A;Ewing AG
通讯作者：
Ewing AG
影响因子：
4.5
作者：
BOND, AM;LUSCOMBE, D;ZOSKI, CG
通讯作者：
ZOSKI, CG
影响因子：
4.7
作者：
CAPELLA, P;GHASEMZADEH, MB;WIGHTMAN, RM
通讯作者：
WIGHTMAN, RM
影响因子：
7.4
作者：
Cui, Hui-Fang;Ye, Jian-Shan;Sheu, Fwu-Shan
通讯作者：
Sheu, Fwu-Shan