High temporal resolution for in vivo monitoring of neurotransmitters in awake epileptic rats using brain microdialysis and capillary electrophoresis with laser-induced fluorescence detection
- 作者：Parrot ; Sandrine ; Sauvinet ; Valé ; rie ; Riban ; Vé ; ronique ; Depaulis ; Antoine ; Renaud ; Bernard ; et. al.
- 关键词：Amino acids ; Catecholamines ; High sampling rate microdialysis ; Awake rats
- 刊名：Journal of Neuroscience Methods
- 出版年：2004
- 期刊代码：33_01650270
- 类别：neu
- 出版时间：December 30, 2004
- 卷：140
- 期：1-2
- 页码：29-38
- 文件大小：197 K
摘要
A method for high temporal resolution monitoring of five neurotransmitters, dopamine (DA), noradrenaline (NA), gamma-aminobutyric acid (GABA), glutamate (Glu), l-aspartate (L-Asp), in freely-moving rats using microdialysis and capillary electrophoresis with laser-induced fluorescence detection (CE-LIFD) was developed. An on-line device, including microdialysis and derivatization with naphthalene-2,3-dicarboxaldehyde, mixes the dialysate with derivatization reagents directly in the collection tube, i.e. with no reactor. Thereafter, collected derivatized samples are analyzed off-line with an automated CE system coupled to a LIFD using a 442nm excitation. The sampling time was limited by the minimal volume required for the analysis by the automated CE system used: neurotransmitters could be determined in 667nl dialysates (940nl after derivatization), i.e. in samples collected every 20s with a flow rate of 2μl/min. The detection limits at the dialysis probe were 3 × 10−9, 1 × 10−9, 1.9 × 10−8, 4.2 × 10−7, 2.1 × 10−7mol/l for DA, NA, GABA, Glu and L-Asp, respectively. The protocol was validated using in vitro/in vivo tests and the performances—repeatability, linearity, characteristics of the probes—were determined.