Glick S D, Dong N, Keller R W, Carlson J N
Department of Pharmacology and Toxicology, Albany Medical College, New York 12208-3479.
J Neurochem. 1994 May;62(5):2017-21. doi: 10.1046/j.1471-4159.1994.62052017.x.
It is common practice in microdialysis studies for probes to be "calibrated" in artificial CSF and in vitro recoveries determined for all substances to be measured in vivo. Dialysate concentrations of such substances are then "corrected" for in vitro recoveries to provide "estimates" of extracellular concentrations. At least for dopamine, in vitro and in vivo recoveries are significantly different and, therefore, an estimate of extracellular dopamine based on correction for in vitro recovery is likely to be erroneous. Generally, however, the relative relationships of such estimates among animals are of interest rather than the "true" extracellular values. Such relationships would be valid to the extent that estimated values are correlated with or predictive of true values. Using the "no net flux" procedure, the present study sought to determine, for both dopamine and its metabolite 3,4-dihydroxyphenylacetic acid (DOPAC), whether in vitro and in vivo recoveries would correlate with each other as well as whether respective estimated and true (no net flux) values of these substances would correlate with each other. Probes (3 mm; BAS/CMed MF-5393), previously calibrated, were lowered into both the nucleus accumbens and striatum of freely moving rats the day before sample collection was begun. In vitro and in vivo recoveries were not significantly correlated (r = 0.1-0.3), for either dopamine or DOPAC. For both dopamine and DOPAC, however, there were significant correlations (r = 0.7-0.8) between estimated and true values. Surprisingly, when using these commercial probes, absolute dialysate levels for both substances were even better correlated (r = 0.9-0.95) with true values.(ABSTRACT TRUNCATED AT 250 WORDS)
在微透析研究中,常见的做法是将探针在人工脑脊液中进行“校准”,并测定所有要在体内测量的物质的体外回收率。然后针对体外回收率对这些物质的透析液浓度进行“校正”,以提供细胞外浓度的“估计值”。至少对于多巴胺来说,体外和体内回收率存在显著差异,因此,基于体外回收率校正得出的细胞外多巴胺估计值可能是错误的。然而,一般来说,动物之间这些估计值的相对关系才是研究的重点,而非“真实的”细胞外值。只要估计值与真实值相关或具有预测性,这种关系就是有效的。本研究采用“无净通量”程序,旨在确定多巴胺及其代谢物3,4 - 二羟基苯乙酸(DOPAC)的体外和体内回收率是否相互关联,以及这些物质各自的估计值与真实值(无净通量)是否相互关联。在开始采集样本的前一天,将预先校准的探针(3毫米;BAS/CMed MF - 5393)插入自由活动大鼠的伏隔核和纹状体。多巴胺和DOPAC的体外和体内回收率均无显著相关性(r = 0.1 - 0.3)。然而,对于多巴胺和DOPAC,估计值与真实值之间均存在显著相关性(r = 0.7 - 0.8)。令人惊讶的是,使用这些商用探针时,两种物质的绝对透析液水平与真实值的相关性甚至更好(r = 0.9 - 0.95)。(摘要截选至250字)