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REGIONAL LEVELS OF GABA IN THE BRAIN: RAPID SEMIAUTOMATED ASSAY AND PREVENTION OF POSTMORTEM INCREASE BY 3‐MERCAPTO‐PROPIONIC ACID
Author(s) -
Heyden J. A. M.,
Korf J.
Publication year - 1978
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1978.tb12448.x
Subject(s) - substantia nigra , chemistry , detection limit , reagent , central nervous system , chromatography , brain tissue , microbiology and biotechnology , biochemistry , biology , endocrinology , anatomy , dopamine , dopaminergic
— 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.