In vivo electrochemical measurements of exogenously applied dopamine in Drosophila melanogaster.
In vivo electrochemical measurements of exogenously applied dopamine in Drosophila melanogaster.
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DOI：
10.1021/ac802297b
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发表时间：
2009-03-01
影响因子：
7.4
通讯作者：
Ewing, Andrew G.
中科院分区：
文献类型：
--
作者：
Makos, Monique A.;Kim, Young-Cho;Han, Kyung-An;Heien, Michael L.;Ewing, Andrew G.
碳纤维微电极与电化学检测联用已被广泛应用于生物胺的分析。为了确定这些胺的功能作用，体内研究主要使用大鼠和小鼠作为模型生物。在这里，我们报告了这些微量分析技术的发展，用于体内电化学检测成年果蝇中枢神经系统(CNS)中的多巴胺。使用三管微吸管注射器外源性施加三种不同浓度的多巴胺，并在多巴胺神经元密集的原脑前内侧区放置圆柱形碳纤维微电极。背景减去快速扫描循环伏安法用于测量飞行中枢神经系统中的多巴胺浓度。野生型果蝇和缺乏功能性多巴胺转运体的富明突变体在清除外源应用的多巴胺方面显示出明显的差异。在可卡因孵育后，野生型果蝇由于多巴胺氧化引起的电流反应比基线显著增加。有趣的是，在相同的条件下，突变果蝇的电流保持不变。这些数据证实了公认的理论，即可卡因阻断了多巴胺转运体的功能，并验证了使用体内电化学方法来监测果蝇对多巴胺的摄取。此外，与钠通道阻滞剂河豚毒素(TTX)孵育后，野生型果蝇的峰值氧化电流显著增加，而fMN突变体的峰值氧化电流没有明显变化。这些数据表明，通过TTX等离子通道影响神经元活动的因素也会影响果蝇体内多巴胺转运体的功能。
Carbon-fiber microelectrodes coupled with electrochemical detection have been used extensively for the analysis of biogenic amines. In order to determine the functional role of these amines, in vivo studies have primarily used rats and mice as model organisms. Here, we report on the development of these microanalytical techniques for in vivo electrochemical detection of dopamine in the adult Drosophila melanogaster central nervous system (CNS). A triple-barrel micropipette injector was used to exogenously apply three different concentrations of dopamine, and a cylindrical carbon-fiber microelectrode was placed in the protocerebral anterior medial brain area where dopamine neurons are densely populated. Background-subtracted fast-scan cyclic voltammetry was used to measure dopamine concentration in the fly CNS. Distinct differences are shown for the clearance of exogenously applied dopamine in the brains of wild type flies versus fumin (fmn) mutants lacking a functional dopamine transporter. The current response due to oxidation of dopamine increased significantly from baseline for wild type flies following cocaine incubation. Interestingly, the current remained unchanged for mutant flies under the same conditions. These data confirm the accepted theory that cocaine blocks dopamine transporter function and validates the use of in vivo electrochemical methods to monitor dopamine uptake in Drosophila. Furthermore, after incubation with tetrodotoxin (TTX), a sodium channel blocker, there was a significant increase in peak oxidation current in the wild type flies; however, the current did not significantly change in the fmn mutant. These data suggest that factors that affect neuronal activity via ion channels such as TTX also influence the function of the dopamine transporter in Drosophila.
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影响因子：
56.9
作者：
EWING, AG;BIGELOW, JC;WIGHTMAN, RM
通讯作者：
WIGHTMAN, RM
影响因子：
4.2
作者：
Heien, MLAV;Phillips, PEM;Wightman, RM
通讯作者：
Wightman, RM
影响因子：
5.3
作者：
Kume, K;Kume, S;Jackson, FR
通讯作者：
Jackson, FR
DOI：
10.1085/jgp.47.5.965
发表时间：
1964-05
期刊：
The Journal of general physiology
影响因子：
--
作者：
NARAHASHI T;MOORE JW;SCOTT WR
通讯作者：
SCOTT WR
影响因子：
3.6
作者：
NASSEL, DR;ELEKES, K
通讯作者：
ELEKES, K