Intracerebral microdialysis study of glutamate reuptake in awake, behaving rats

The central nervous system has high-affinity uptake systems for the clearance of amino acid transmitters. These systems are found in both neurons and astrocytes. Previous studies have shown that the uptake of amino acid transmitters by astrocytes in culture can be modulated by adrenergic agents. The objectives of this study were to develop a methodology that evaluates the brain's reuptake capacity for glutamate in awake, behaving animals and to determine whether glutamate reuptake is under alpha-adrenergic regulation in the intact central nervous system. Male Sprague-Dawley rats weighing 250-450 g were used in this study. The extraction fraction of L-[3H]glutamate with [14C]mannitol as a reference was measured. The cortical extraction fraction of L-[3H]glutamate corrected for [14C]mannitol (EL-glu) reaches steady state rapidly and is both stable and repeatable. EL-glu is a measure of L-glutamate reuptake and not metabolism. EL-glu is decreased in a dose-dependent manner by the addition of the glutamate reuptake blocker D,L-threo-beta-hydroxyaspartic acid or unlabeled L- glutamate. In addition, EL-glu is increased in a dose-dependent manner by the alpha1-adrenergic agonist phenylephrine, and this increase is blocked by the alpha-adrenergic antagonist phentolamine.