Quantitative measurement of transmitters in individual vesicles in the cytoplasm of single cells with nanotip electrodes.
Quantitative measurement of transmitters in individual vesicles in the cytoplasm of single cells with nanotip electrodes.
复制标题
DOI：
10.1002/anie.201504839
复制
发表时间：
2015-10-05
期刊：
影响因子：
--
通讯作者：
Ewing AG
中科院分区：
文献类型：
--
作者：
Li X;Majdi S;Dunevall J;Fathali H;Ewing AG
囊泡递质含量的定量对于研究疾病中神经传递和功能障碍的机制非常重要，但测量单个囊泡阿托升体积中的少量神经递质含量非常困难，尤其是在细胞环境中。我们介绍了一种新方法，细胞内囊泡电化学细胞术。当这些囊泡在活细胞内的电极上裂解时，使用纳米尖端锥形碳纤维微电极以电化学方式测量单个 PC12 细胞中单个纳米级囊泡中电活性神经递质的总含量。结果表明，在胞吐作用过程中，仅释放了神经递质量子含量的一小部分。这些数据支持了一个有趣的假设，即在正常的胞吐过程中，囊泡不会完全打开，导致囊泡内容物不完全排出。
Quantification of vesicular transmitter contents is important for studying the mechanisms of neurotransmission and malfunction in disease and yet it is incredibly difficult to measure the small contents of neurotransmitters in the attoliter volume of a single vesicle, especially in the cell environment. We introduce a novel method, intracellular vesicle electrochemical cytometry. A nano-tip conical carbon fiber microelectrode is used to electrochemically measure the total contents of electroactive neurotransmitters from individual nanoscale vesicles in single PC12 cells as these vesicles lyse on the electrode inside the living cell. The results demonstrate that only a fraction of quantal contents of neurotransmitter is released during exocytosis. These data support the intriguing hypothesis that the vesicle does not open all the way during the normal exocytosis process, resulting in incomplete expulsion of the vesicular contents.