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Effect of potassium and N-methyl-D-aspartate on the aspartate aminotransferase activity in cultured cerebellar granule cells

✍ Scribed by J. Morán; M. Rivera-Gaxiola

Publisher
John Wiley and Sons
Year
1992
Tongue
English
Weight
892 KB
Volume
33
Category
Article
ISSN
0360-4012

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✦ Synopsis

The effect of potassium depolarization and N-methyl-D-aspartate (NMDA) on the activity of aspartate aminotransferase (AAT; EC 2.6.1.1), an enzyme suggested to be involved in neurotransmitter glutamate synthesis, was studied in cultured cerebellar granule neurons. Both KCI and NMDA increased AAT activity in a dose-dependent manner. When cells were treated 48-72 hr with 40 mM KCl or 150 pM NMDA the AAT was enhanced about 65-75%. The EC,, for NMDA and KCI were 25 pM and 17 mM, respectively. The effect of NMDA and KCI was specific for AAT without affecting the activity of other enzymes like lactate dehydrogenase or protein content and it was observed only in granule cells but not in astrocytes or cortical neurons. The effect of KCI was not mediated by an activation of excitatory amino acid receptors and was Ca+ +-dependent. The effect of NMDA was completely blocked by Mg++ and NMDA antagonists. The increase of AAT induced by AAT and KCl was blocked by cycloheximide and actinomycin D, suggesting an involvement of de novo synthesis of proteins and RNA. Kainic acid and quinolinic acid were also effective in increasing the AAT activity. The action of kainate was less effective than that of NMDA and it was observed only at relatively low concentrations (10 pM). Quinolinic acid raised the activity of AAT about 45% at a concentration of 500 pM. Other non-NMDA agonists did not modify the AAT activity. From these findings we can conclude that NMDA and KCl exert a trophic action on cerebellar granular neurons. These conditions could be a simulation of the presynaptic activity required during development to induce survival and a biochemical differentiation, which we evaluated using AAT as a marker enzyme.

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