Analysis of extracellular gamma-aminobutyric acid, glutamate and aspartate in cat visual cortex by in vivo microdialysis and capillary electrophoresis-laser induced fluorescence detection.
To investigate the influence of a partial sensory deprivation on the extracellular concentration of amino acid neurotransmitters in cat visual cortex, a capillary electrophoresis method was developed for the quantification of gamma-aminobutyric acid (GABA), glutamate (Glu) and aspartate (Asp) in in vivo microdialysis samples of cat brain. Microdialysis samples from different regions of area 17 were obtained every 15-min using CMA 12 2-mm probes perfused with synthetic cerebrospinal fluid and derivatized using fluorescein isothiocyanate (FITC). Laser-induced fluorescence (LIF) detection was employed. Good selectivity was obtained with a borate buffer (20 mM, pH 10.25). The whole procedure, including the washing step takes only 15 min. The conditions for derivatization and separation were optimized. The parameters for validation such as linearity, precision and detection limit are also reported. The results are consistent with those of HPLC but, as the sample volumes needed are only 1--5 nl, a much better time resolution can be obtained.
Laboratory for Neuroendocrinology and Immunological Biotechnology, Zoological Institute, Katholieke Universiteit Leuven, Leuven, Belgium.
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Li
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Vandenbussche
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Arckens
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Vandesande
D000818:Animals; D001224:Aspartic Acid / Q000032:analysis; D002415:Cats; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D018698:Glutamic Acid / Q000032:analysis; D007834:Lasers; D017551:Microdialysis / Q000295:instrumentation / Q000379:methods*; D014793:Visual Cortex / Q000737:chemistry*; D005680:gamma-Aminobutyric Acid / Q000032:analysis
D005456:Fluorescent Dyes; D001224:Aspartic Acid; D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid; D016650:Fluorescein-5-isothiocyanate