Characterization of dopamine transport in crude synaptosomes prepared from rat medial prefrontal cortex
Characterization of dopamine transport in crude synaptosomes prepared from rat medial prefrontal cortex
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DOI：
10.1016/j.jneumeth.2004.02.022
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发表时间：
2004-08-30
影响因子：
3
通讯作者：
Steketee, JD
中科院分区：
文献类型：
--
作者：
Williams, JA;Steketee, JD
越来越多的证据表明，多巴胺（DA）的摄取到mesocortical神经元可能是通过调节机制，显着不同的黑质纹状体或mesocorticbens系统中观察到的。本研究旨在建立一种快速、灵敏的单一动物内侧前额叶皮质（mPFC）粗突触体DA摄取测定方法。DA摄取到mPFC是饱和的，线性相对于蛋白质浓度，时间依赖性，和敏感的单胺转运抑制剂的影响。饱和度分析显示，DA转运在mPFC中的Km和V-max值分别约为60 nM和6.5 pmol/min mg蛋白。与GBR 12909、氟西汀（FLX）和可卡因（COC）相比，mPFC中显著量的DA摄取对nisoxetine的抑制更敏感，表明去甲肾上腺素转运蛋白（NET）在该区域内DA的清除中起重要作用。所描述的测定条件将可用于检查从单个动物获得的特定脑区域内的DA摄取。(C)2004 Elsevier B. V.保留所有权利。
Accumulating evidence suggests that dopamine (DA) uptake into mesocortical neurons may be regulated through mechanisms that are markedly different from those observed in nigrostriatal or mesoaccumbens systems. The current studies were conducted to develop a rapid and sensitive DA uptake assay in crude synaptosomes prepared from the medial prefrontal cortex (mPFC) of a single animal. Uptake of DA into the mPFC was saturable, linear with respect to protein concentration, time dependent, and sensitive to the effects of monoamine transport inhibitors. Saturation analysis revealed the K-m and V-max values for DA transport in the mPFC were approximately 60 nM and 6.5 pmol/min mg protein, respectively. A significant amount of DA uptake in the mPFC was more sensitive to inhibition by nisoxetine compared to GBR12909, fluoxetine (FLX), and cocaine (COC), suggesting the norepinephrine transporter (NET) plays an important role in the clearance of DA within this region. The described assay conditions would be useful in examining DA uptake within specific brain regions obtained from a single animal. (C) 2004 Elsevier B.V. All rights reserved.