Ciriacks Chanda M, Bowser Michael T
Department of Chemistry, University of Minnesota, 207 Pleasant Street SE, Minneapolis, Minnesota 55455, USA.
Anal Chem. 2004 Nov 15;76(22):6582-7. doi: 10.1021/ac0490651.
D-Serine was detected in dialysate collected from the rat striatum using an online microdialysis-CE-LIF instrument. Dialysate can be analyzed every 12.5 s using the online instrument, giving much better temporal resolution than previously possible for D-serine. Basal concentrations of D-serine (8 +/- 2 microM), glutamate (0.8 +/- 0.2 microM), GABA (0.11 +/- 0.04 microM), and L-serine (23 +/- 4 microM) were measured. Increases in the concentrations of these neurochemicals induced by the introduction of high-K+ aCSF were quantitated. Notably, an increase in D-serine concentration in response to high-K+ aCSF was observed for the first time. The identity of the D-serine peak was confirmed unambiguously using D-amino acid oxidase to selectively remove D-serine from a dialysate sample. The microdialysis-CE-LIF instrument was able to monitor this enzymatic reaction as it proceeded over a period of 60 min, demonstrating that online microdialysis-CE-LIF is not only useful in monitoring in vivo dynamics but can also be used to monitor other chemical systems.
使用在线微透析-毛细管电泳-激光诱导荧光(CE-LIF)仪器在从大鼠纹状体收集的透析液中检测到了D-丝氨酸。使用该在线仪器可每12.5秒分析一次透析液,这比以前检测D-丝氨酸时能获得更好的时间分辨率。测量了D-丝氨酸(8±2微摩尔)、谷氨酸(0.8±0.2微摩尔)、γ-氨基丁酸(GABA,0.11±0.04微摩尔)和L-丝氨酸(23±4微摩尔)的基础浓度。对引入高钾人工脑脊液(aCSF)后这些神经化学物质浓度的增加进行了定量。值得注意的是,首次观察到高钾aCSF引起D-丝氨酸浓度升高。使用D-氨基酸氧化酶从透析液样品中选择性去除D-丝氨酸,明确证实了D-丝氨酸峰的身份。微透析-CE-LIF仪器能够在60分钟的时间内监测该酶促反应的进行过程,表明在线微透析-CE-LIF不仅可用于监测体内动态,还可用于监测其他化学系统。