REGIONAL LEVELS OF GABA IN THE BRAIN: RAPID SEMIAUTOMATED ASSAY AND PREVENTION OF POSTMORTEM INCREASE BY 3-MERCAPTO-PROPIONIC ACID |
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Authors: | J. A. M. van der Heyden J. Korf |
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Affiliation: | Department of Biological Psychiatry, Psychiatric University Clinic, Oostersingel 59, Groningen, The Netherlands |
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Abstract: | Abstract— A simple and rapid semiautomated assay for GABA in central nervous tissue is described. The method is based on a simple manual procedure of isolating GABA from tissue extracts on small CM Sepharose Cl 6B columns, followed by an automated fluorimetric detection (continuous flow system) with o -phthalaldehyde (OPA) and β-mercapto-ethanol (β-ME) at an alkaline pH. GABA is separated from other compounds that fluoresce in our detection system. By using low concentration of OPA and β-ME and allowing only a short reaction time with these reagents, the detection is specific towards GABA. The detection limit of the assay is 1 nmol. A procedure is described for the prevention of postmortem GABA increase in rat and mouse brain by intravenous injection of 3-mercapto-propionic acid (1.2 nmol/kg) 2min before decapitating the animal. This treatment and microwave irradiation result in similar GABA levels in mouse brain and substantia nigra tissue from rat brain. We found a great conformity in regional GABA levels in the rat and the mouse brain. |
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