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利用胶束电动毛细管色谱-电化学检测法测定果蝇微小脑区中的生物胺。

Biogenic amines in microdissected brain regions of Drosophila melanogaster measured with micellar electrokinetic capillary chromatography-electrochemical detection.

机构信息

Department of Chemistry, The Pennsylvania State University, 125 Chemistry Building, University Park, Pennsylvania 16802, USA.

出版信息

Anal Chem. 2010 Sep 15;82(18):7729-35. doi: 10.1021/ac101603d.

Abstract

Micellar electrokinetic chromatography with electrochemical detection has been used to quantify biogenic amines in microdissected Drosophila melanogaster brains and brain regions. The effects of pigment from the relatively large fly eyes on the separation have been examined to find that the red pigment from the compound eye masks much of the signal from biogenic amines. The brains of white mutant flies, which have characteristically low pigment in the eyes, have a significantly simplified separation profile in comparison to the red-eyed, wild-type, Canton S fly. Yet, the white mutant flies were found to have significantly less amounts of dopamine, l-3,4-dihydroxyphenylalanine (L-DOPA), salsolinol, and N-acetyltyramine in their dissected brains when compared to dissected brains of Canton S flies. In addition, significant variation has been observed in the dissected brains between individual flies that might be related to changes in neurotransmitter turnover. The transgenic GFP fly line (TH-GFP), for which the overall profile of biogenic amines is not found to be significantly different from Canton S, can be used to visualize the location of dopamine neurons. Biogenic amines were then quantified in three brain regions observed to have dopamine levels, the central brain, optic lobes, and posterior superiormedial protocerebrum (PPM1) region.

摘要

胶束电动色谱-电化学检测已被用于定量分析微切割黑腹果蝇大脑和脑区中的生物胺。考察了来自相对较大的果蝇眼睛的色素对分离的影响,发现复眼的红色色素掩盖了生物胺的大部分信号。与红眼睛的野生型 Canton S 果蝇相比,具有特征性低色素的白眼突变体果蝇的分离谱显著简化。然而,与 Canton S 果蝇的脑部分离物相比,白眼突变体果蝇的脑部分离物中多巴胺、l-3,4-二羟基苯丙氨酸 (L-DOPA)、salsolinol 和 N-乙酰基酪氨酸的含量明显减少。此外,在不同个体的脑部分离物中观察到明显的变化,这可能与神经递质周转率的变化有关。TH-GFP 转基因 GFP 蝇系(TH-GFP)的生物胺总谱与 Canton S 没有显著差异,可用于可视化多巴胺神经元的位置。然后在三个被观察到具有多巴胺水平的脑区(中央脑、视叶和后上侧脑前脑区 (PPM1))中定量分析了生物胺。

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