In situ temporal detection of dopamine exocytosis from L-dopa-incubated MN9D cells using microelectrode array-integrated biochip
In situ temporal detection of dopamine exocytosis from L-dopa-incubated MN9D cells using microelectrode array-integrated biochip
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DOI：
10.1016/j.snb.2005.10.042
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发表时间：
2006-06-26
影响因子：
8.4
通讯作者：
Sheu, Fwu-Shan
中科院分区：
文献类型：
--
作者：
Cui, Hui-Fang;Ye, Jian-Shan;Sheu, Fwu-Shan
多巴胺（DA）是一种重要的神经递质，在许多神经系统疾病中起着非常重要的作用。一个微电极阵列集成的生物芯片已在这里表示为一个方便的装置，用于在原位时间检测多巴胺能细胞的DA胞吐。生物芯片是硅基的，并且在硅板的1 mm中心上间隔开5 x 5阵列的Au盘微电极。MN 9D，小鼠中脑多巴胺能细胞系，已经生长在生物芯片室的表面上。用电流法检测MN 9D细胞DA的胞吐作用。DA在生物芯片微电极上的安培检测限为0.06 ~ 0.21 RM（S/N = 3），K+诱导的MN 9D细胞DA胞吐水平检测不到。相反，MN 9D细胞与多巴胺前体L-dopa孵育后，电流法检测到K+诱导的DA胞吐。K+诱导的DA释放是浓度依赖性的，并且似乎在最大细胞外DA浓度281 +/- 137 nM（平均S.E.）高效液相色谱分析表明，L-多巴诱导的MN 9D细胞K+刺激的胞吐主要含有DA，DA：NE：L-多巴：5-羟色胺的重量比为1.00：0.28：0.06：0.14。这些结果表明MN 9D细胞具有多巴胺能细胞的典型机械系统，包括L-芳香酸脱羧酶、囊泡单胺转运体、电压敏感性Na+通道和电压敏感性钙通道。(c)2005 Elsevier B. V.保留所有权利。
Dopamine (DA) is an important neurotransmitter, playing a very important role in many neurological disorders. A microelectrode array-integrated biochip has been represented here as a convenient device for in situ temporal detection of DA exocytosis from dopaminergic cells. The biochip is silicon-based and a 5 x 5 array of Au disk microelectrodes is spaced on the 1 mm center of the silicon plate. MN9D, a mouse mesencephalic dopaminergic cell line, has been grown on the surface of the biochip chamber. DA exocytosis from the chip-grown MN9D cells was detected by using amperometry. With the amperometric detection limit of DA at the biochip microelectrodes ranging from 0.06 to 0.21 RM (S/N = 3), the level of K+-induced DA exocytosis from MN9D cells was undetectable. In contrast, after MN9D cells were incubated with L-dopa, a DA precursor, K+-induced DA exocytosis was temporally detected by amperometry. The K+-induced DA release is concentration-dependent and appears to be saturated at the maximum extracellular DA concentration of 281 +/- 137 nM (mean S.E.) for 8000 viable MN9D cells, when the extracellular K+ concentration increases to 35 mM. High-performance liquid chromatography demonstrates that the K+-stimulated exocytosis from L-dopa-incubated MN9D cells mainly contains DA, and the weight ratio of DA:NE:L-dopa: serotonin is 1.00:0.28:0.06:0.14. These results suggest that MN9D cell has a typical machinery system of a dopaminergic cell, including L-aromatic acid decarboxylase, vesicular monoamine transporter, voltage-sensitive Na+ channels, and voltage-sensitive calcium channels. (c) 2005 Elsevier B.V. All rights reserved.