Abstract— To obtain evidence of the site of conversion of [U‐14C]glucose into glutamate and related amino acids of the brain, a mixture of [U‐14C]glucose and [3H]glutamate was injected subcutaneously into rats. [3H]Glutamate gave rise to several 3H‐labelled amino acids in rat liver and blood; only 3H‐labelled glutamate, glutamine or γ‐aminobutyrate were found in the brain. The specific radioactivity of [3H]glutamine in the brain was higher than that of [3H]glutamate indicating the entry of [3H]glutamate mainly in the ‘small glutamate compartment’. The 14C‐labelling pattern of amino acids in the brain and liver after injection of [U‐14C]glucose was similar to that previously reported (Gaitondeet al., 1965). The specific radioactivity of [14C]glutamine in the blood and liver after injection of both precursors was greater than that of glutamate between 10 and 60 min after the injection of the precursors. The extent of labelling of alanine and aspartate was greater than that of other amino acids in the blood after injection of [U‐14C]glucose. There was no labelling of brain protein with [3H]glutamate during the 10 min period, but significant label was found at 30 and 60 min. The highest relative incorporation of [14C]glutamate and [14C]aspartate in rat brain protein was observed at 5 min after the injection of [U‐14C]glucose. The results have been discussed in the context of transport of glutamine synthesized in the brain and the site of metabolism of [U‐14C]glucose in the brain.