Steady-state in vivo glutamate dehydrogenase activity in rat brain measured by 15N NMR

Abstract

The in vivo activity of glutamate dehydrogenase (GDH) in the direction of reductive amination was measured in rat brain at steady-state concentrations of brain ammonia and glutamate after intravenous infusion of the substrate 15NH4+. The in vivo rate was determined from the steady-state fractional 15N enrichment of brain ammonia, measured by selective observation of 15NH4+ protons in brain extract by 1H-15N heteronuclear multiple-quantum coherence transfer NMR, and the rate of increase of brain [15N]glutamate and [2-15N]glutamine measured by 15N NMR. The in vivo GDH activity was 0.76-1.17 mumol/h/g, and 1.1-1.2 mumol/h/g at 1.0 +/- 0.17 mumol/g. Comparison of the observed in vivo GDH activity with the in vivo rates of glutamine synthesis and of phosphate-activated glutaminase suggests that, under mild hyperammonemia, GDH-catalyzed de novo synthesis can provide a minimum of 19% of the glutamate pool that is recycled from neurons to astrocytes through the glutamate-glutamine cycle.