Outflow and overflow of picogram levels of endogenous dopamine and DOPAC from rat striatal slices: improved methodology for studies of stimulus-evoked release and metabolism.
Outflow and overflow of picogram levels of endogenous dopamine and DOPAC from rat striatal slices: improved methodology for studies of stimulus-evoked release and metabolism.
复制标题
大鼠纹状体切片中皮克级内源性多巴胺和 DOPAC 的流出和溢出：刺激诱发释放和代谢研究的改进方法。
DOI：
10.1016/0165-0270(89)90119-2
复制
发表时间：
1989
影响因子：
3
通讯作者：
Zahniser,NR
中科院分区：
文献类型：
--
作者：
Gerhardt,GA;Dwoskin,LP;Zahniser,NR
本文报道了一种改进的高效液相色谱-电化学库仑检测法（HPLC-EC）测定大鼠纹状体脑片灌流液中内源性多巴胺（DA）和3，4-二羟基苯乙酸（DOPAC）的方法。在加入少量溶解的抗坏血酸氧化酶后，将超灌注液直接注入HPLC-EC系统。DA和DOPAC均在3-5分钟内分离和定量。每小时可分析12至15份样品，每种化合物的标称检测限为1.0 pg/注射或10-20 pg/ml灌流液。本方法被用来研究DA和DOPAC流出和溢出的变化，在灌流液中的单个纹状体切片电场刺激后，无论是在存在和不存在的儿茶酚胺摄取抑制剂诺米芬新。1分钟的灌流收集的研究清楚地表明，电场刺激产生了潜在的增加DOPAC相比DA。常规的灵敏度和样品通量的方法允许研究的流出和溢出的DA和DOPAC，以及研究involvingtime-dependentoverflow这些化合物。
An improved method for the rapid and sensitive determination of endogenous dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) in superfusates of single rat striatal slices, using high-performance liquid chromatography (HPLC) coupled with ‘coulometric’ electrochemical detection (EC), is described. Superfusates are directly injected into an HPLC-EC system following addition of a small aliquot of solubilized ascorbate-oxidase. DA and DOPAC are both separated and quantitated in 3–5 min. Twelve to 15 samples can be analyzed each hour with a nominal detection limit of 1.0 pg per injection for each compound or 10–20 pg/ml of superfusate. The present method was used to study changes in DA and DOPAC outflow and overflow in superfusates of single striatal slices following electrical field stimulation, both in the absence and presence of the catecholamine uptake inhibitor nomifensine. Studies of 1 min superfusate collections clearly showed that electrical field stimulation produced a latent increase in DOPAC as compared to DA. The routine sensitivity and sample throughput of the method allows for studies of both outflow and overflow of DA and DOPAC, as well as studies involvingtime-dependentoverflow of these compounds.