使用微石墨化金刚石多电极阵列同时检测 PC12 细胞的量子释放。

Simultaneous multisite detection of quantal release from PC12 cells using micro graphitic-diamond multi electrode arrays.

机构信息

Department of Drug and Science Technology, NIS Inter-departmental Centre, University of Torino, Corso Raffaello 30, 10125 Torino, Italy.

出版信息

Biophys Chem. 2019 Oct;253:106241. doi: 10.1016/j.bpc.2019.106241. Epub 2019 Jul 31.

DOI:10.1016/j.bpc.2019.106241

PMID:31398633

Abstract

Micro graphitic - diamond - multi electrode arrays (μG-D-MEAs) are suitable for measuring multisite quantal dopamine (DA) release from PC12 cells. Following cell stimulation with high extracellular KCl and electrode polarization at +650 mV, amperometric spikes are detected with a mean frequency of 0.60 ± 0.16 Hz. In each recording, simultaneous detection of secretory events is occurred in approximately 50% of the electrodes. Kinetic spike parameters and background noise are preserved among the different electrodes. Comparing the amperometric spikes recorder under control conditions with those recorders from PC12 cells previously incubated for 30 min with the dopamine precursor Levodopa (L-DOPA, 20 μM) it appears that the quantal size of amperometric spikes is increased by 250% and the half-time width (t) by over 120%. On the contrary, L-DOPA has no effect on the frequency of secretory events. Overall, these data demonstrate that the μG-D-MEAs represent a reliable bio-sensor to simultaneously monitor quantal exocytotic events from different cells and in perspective can be exploited as a drug-screening tool.

摘要

微石墨 - 金刚石 - 多电极阵列（μG-D-MEAs）适用于测量 PC12 细胞中多部位量子多巴胺（DA）的释放。在用高细胞外 KCl 刺激细胞后，将电极极化至+650 mV，以 0.60 ± 0.16 Hz 的平均频率检测到安培计尖峰。在每次记录中，约有 50%的电极同时检测到分泌事件。不同电极之间保留了动力学尖峰参数和背景噪声。将对照条件下的安培计尖峰记录器与先前用多巴胺前体左旋多巴（L-DOPA，20 μM）孵育 30 分钟的 PC12 细胞的记录器进行比较，似乎安培计尖峰的量子大小增加了 250%，半衰期（t）超过 120%。相反，L-DOPA 对分泌事件的频率没有影响。总的来说，这些数据表明 μG-D-MEAs 代表了一种可靠的生物传感器，可以同时监测来自不同细胞的量子胞吐事件，并且可以作为药物筛选工具加以利用。