Using In Vivo Electrochemistry to Study the Physiological Effects of Cocaine and Other Stimulants on the Drosophila melanogaster Dopamine Transporter.
Using In Vivo Electrochemistry to Study the Physiological Effects of Cocaine and Other Stimulants on the Drosophila melanogaster Dopamine Transporter.
复制标题
利用体内电化学研究可卡因和其他兴奋剂对果蝇多巴胺转运蛋白的生理影响。
DOI：
10.1021/cn900017w
复制
发表时间：
2010-01-20
影响因子：
5
通讯作者：
Ewing, Andrew G.
中科院分区：
文献类型：
--
作者：
Makos, Monique A.;Han, Kyung-An;Heien, Michael L.;Ewing, Andrew G.
多巴胺神经传递被认为在药物滥用的成瘾强化机制中起关键作用。电化学技术已被广泛用于监测包括大鼠、小鼠和灵长类动物在内的模式生物脑中的体内多巴胺变化。在这里，我们研究了几种兴奋剂对多巴胺清除率的影响，使用最近开发的微量分析工具，在体内电化学测量多巴胺在果蝇中枢神经系统。一个圆柱形的碳纤维微电极被放置在果蝇大脑的前脑前内侧区（多巴胺神经元密集的区域），同时定位一个微量移液管注射器以外源性地施加多巴胺。背景扣除快速扫描循环伏安法进行定量多巴胺浓度的变化，在成年苍蝇的大脑。清除外源应用多巴胺显着降低了野生型苍蝇的前大脑前内侧区可卡因，苯丙胺，甲基苯丙胺，或哌甲酯治疗后。与此相反，多巴胺摄取保持不变时，采用相同的治疗缺乏功能性多巴胺转运蛋白的fumin突变苍蝇。我们的体内结果支持体外结合亲和力研究，预测这四种兴奋剂有效地阻止正常的果蝇多巴胺转运蛋白功能。此外，我们发现10 μ M的生理可卡因浓度足以显著改变果蝇中枢神经系统中多巴胺转运蛋白的摄取。综上所述，这些数据表明，果蝇大脑中的多巴胺摄取减少了精神兴奋剂在哺乳动物中观察到的。这验证了果蝇作为未来研究人类药物成瘾的细胞和分子机制的模型系统的使用。
Dopamine neurotransmission is thought to play a critical role in addiction reinforcing mechanisms of drugs of abuse. Electrochemical techniques have been employed extensively for monitoring in vivo dopamine changes in the brains of model organisms including rats, mice, and primates. Here, we investigated the effects of several stimulants on dopamine clearance using recently developed microanalytical tools for in vivo electrochemical measurements of dopamine in the central nervous system of Drosophila melanogaster. A cylindrical carbon-fiber microelectrode was placed in the protocerebral anterior medial region of the Drosophila brain (an area dense with dopamine neurons) while a micropipette injector was positioned to exogenously apply dopamine. Background-subtracted fast-scan cyclic voltammetry was carried out to quantify changes in dopamine concentration in the adult fly brain. Clearance of exogenously applied dopamine was significantly decreased in the protocerebral anterior medial area of the wild-type fly following treatment with cocaine, amphetamine, methamphetamine, or methylphenidate. In contrast, dopamine uptake remained unchanged when identical treatments were employed in fumin mutant flies that lack functional dopamine transporters. Our in vivo results support in vitro binding affinity studies that predict these four stimulants effectively block normal Drosophila dopamine transporter function. Furthermore, we found 10 μM to be a sufficient physiological cocaine concentration to significantly alter dopamine transporter uptake in the Drosophila central nervous system. Taken together, these data indicate dopamine uptake in the Drosophila brain is decreased by psychostimulants as observed in mammals. This validates the use of Drosophila as a model system for future studies into the cellular and molecular mechanisms underlying drug addiction in humans.
登录
查看更多内容
影响因子：
4.7
作者：
EWING, AG;WIGHTMAN, RM
通讯作者：
WIGHTMAN, RM
影响因子：
2.7
作者：
BALSTER, RL;SCHUSTER, CR
通讯作者：
SCHUSTER, CR
影响因子：
4.2
作者：
Heien, MLAV;Phillips, PEM;Wightman, RM
通讯作者：
Wightman, RM
影响因子：
56.9
作者：
EWING, AG;BIGELOW, JC;WIGHTMAN, RM
通讯作者：
WIGHTMAN, RM
影响因子：
15.9
作者：
KUHAR, MJ;RITZ, MC;BOJA, JW
通讯作者：
BOJA, JW