Previous studies have shown that the glutamate͞glutamine (Glu͞ Gln) neurotransmitter cycle and neuronal glucose oxidation are proportional (1:1), with increasing neuronal activity above isoelectricity. GABA, a product of Glu metabolism, is synthesized from astroglial Gln and contributes to total Glu͞Gln neurotransmitter cycling, although the fraction contributed by GABA is unknown. In the present study, we used 13 C NMR spectroscopy together with i.v. infusions of [1,6-13 C2]glucose and [2-13 C]acetate to separately determine rates of Glu͞Gln and GABA͞Gln cycling and their respective tricarboxylic acid cycles in the rat cortex under conditions of halothane anesthesia and pentobarbital-induced isoelectricity. Under 1% halothane anesthesia, GABA͞Gln cycle flux comprised 23% of total (Glu plus GABA) neurotransmitter cycling and 18% of total neuronal tricarboxylic acid cycle flux. In isoelectric cortex, glucose oxidation was reduced >3-fold in glutamatergic and GABAergic neurons, and neurotransmitter cycling was below detection. Hence, in both cell types, the primary energetic costs are associated with neurotransmission, which increase together as cortical activity is increased. The contribution of GABAergic neurons and inhibition to cortical energy metabolism has broad implications for the interpretation of functional imaging signals.GABAergic neurons ͉ glutamatergic neurons ͉ magnetic resonance spectroscopy ͉ neuronal-glial interactions G lutamate (Glu) and GABA are the major excitatory and inhibitory neurotransmitters in the mature cerebral cortex and together account for Ϸ90% of total cortical synapses (1). Excitatory synapses outnumber inhibitory synapses Ϸ5:1 (2), suggesting that excitation plays an energetically dominant role in the cortex. The energetic cost of GABAergic neurotransmission remains an open question, however, because current methods used to assess cortical activity are based on changes in local blood f low or metabolism (e.g., glucose or oxygen consumption), which cannot differentiate glutamatergic from GABAergic neurons. Thus, the interpretation of changes in cortical activity in terms of the energetics of excitation and inhibition requires other methods that are sensitive to the synthesis of these neurotransmitters.Glutamatergic neurotransmitter cycling flux and energy consumption have been reported for rat and human cortex by using 13 C NMR spectroscopy during the i.v. infusion of 13 C-labeled glucose (3-10). These studies show that the Glu͞glutamine (Gln) cycling flux (V cyc ) is substantial, from Ϸ30% to 42% of neuronal tricarboxylic acid (TCA) cycle flux in anesthetized rats (V TCAn ) (4, 5, 8) to Ϸ38-50% of V TCAn in resting awake rat and human cortex (7, 9, 10). In the cortex of anesthetized rats, changes in V cyc and V TCAn are proportional (Ϸ1:1) over a large cortical activity range above isoelectricity (4,8).The determination of V cyc from Gln and Glu 13 C turnover, using [1-13 C]glucose as tracer, includes contributions from GABA as well as Glu (4,5,11). This is because Gln is a com...