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COMPARTMENTATION OF CITRIC ACID CYCLE METABOLISM IN BRAIN: EFFECT OF AMINOOXYACETIC ACID, OUABAIN AND Ca 2+ ON THE LABELLING OF GLUTAMATE, GLUTAMINE, ASPARTATE AND GABA BY [1‐ 14 C]ACETATE, [U‐ 14 C]GLUTAMATE AND [U‐ 14 C] ASPARTATE 1
Author(s) -
Berl S.,
Clarke D. D.,
Nicklas W. J.
Publication year - 1970
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.1471-4159.1970.tb02253.x
Subject(s) - aminooxyacetic acid , glutamine , glutamate receptor , citric acid cycle , biochemistry , ouabain , glutaminase , glutamate dehydrogenase , metabolism , glutamic acid , amino acid , chemistry , biology , enzyme , sodium , receptor , organic chemistry
—(1) The effects of aminooxyacetic acid, ouabain and Ca 2+ on the compartmentation of amino acid metabolism have been studied in slices of brain incubated with sodium‐[1‐ 14 C]acetate, l‐[U‐ 14 C]glutamate and l‐[U‐ 14 C]aspartate as tracer metabolites. (2) Aminooxyacetic acid (10 ‐3 m) inhibited the labelling of aspartate from [ 14 C]acetate and [ 14 C]glutamate, as well as the incorporation of label from [ 14 C]aspartate into glutamate and glutamine. It also inhibited the labelling of GABA from all three radioactive precursors, as would be anticipated if there was inhibition of several transaminases as well as glutamate decarboxylase. The RSA of glutamine labelled from [1‐ 14 C]acetate was increased. This finding indicated that the glutamate pool which is utilized for glutamine formation is associated with glutamate dehydrogenase, and this enzyme appears to be related to the ‘synthetic tricarboxylic acid cycle’. AOAA exerted its major inhibitory effects on the citric acid‘energy cycle’with which transaminases are associated. (3) Ouabain (10 ‐5 m) inhibited the labelling of glutamine to a much greater extent than the labelling of glutamate from [1‐ 14 C]acetate. It also caused leakage of amino acids from the tissue into the medium. Its effect on the glutamate–glutamine system was interpreted to be a selective inhibition of the 'synthetic’citric acid cycle. (4) The omission of Ca 2+ from the incubation medium was associated with formation of glutamine with RSA less than 1·0 when labelled from [U‐ 14 C]glutamate, [U‐ 14 C]aspartate and lower than normal when labelled from [1‐ 14 C]acetate.