Release Of Amino Acid Neurotransmitters Research Articles

GM1 ganglioside inhibits ischemic release of amino acid neurotransmitters from rat cortex.

The effects of a 20 min period of four-vessel occlusion cerebral ischemia followed by reperfusion on glutamate, aspartate, GABA and glycine efflux from the rat cerebral cortex were studied using a cortical cup technique. Cerebral ischemia increased amino acid concentrations in the cortical superfusates. When the cerebral cortices were exposed to topically applied GM1 ganglioside (50 microM) for 40 min prior to ischemia, the evoked efflux of all four amino acids was significantly inhibited. GM1 (1 microM) failed to inhibit amino acid release. The results support the concept that gangliosides have a cerebroprotective action.

Effect of α2-adrenergic drugs dexmedetomidine and atipamezole on extracellular amino acid levels in vivo

α 2-Adrenoceptors are known to be involved in a variety of physiological functions and pathological conditions, including epilepsy and the extent of excitotoxin-induced cell death. In this study we evaluated whether selective α 2-adrenergic drugs can modulate the release of neurotransmitter amino acids. The effect of the α 2-adrenoceptor agonist dexmedetomidine (5 μg/kg, s.c.) and the α 2-adrenoceptor antagonist atipamezole (0.1 mg/kg and 1 mg/kg, s.c.) on the release of extracellular glutamate, aspartate and γ-aminobutyric acid (GABA) was studied with microdialysis in the hippocampus of freely moving rats under basal and K +-evoked conditions. Atipamezole (1 mg/kg) decreased K +-evoked glutamate efflux by 30% compared to the control group ( P < 0.05) but did not affect significantly the effluxes of aspartate and GABA. Dexmedetomidine and the lower dose of atipamezole (0.1 mg/kg) did not significantly alter the evoked overflow of amino acids. The results suggest that α 2-adrenergic drugs have only modest effects on the K +-stimulated overflow of extracellular neurotransmitter amino acids in rat hippocampus.

A comparison between the stimulated and paroxysmal release of endogenous amino acids from rat cerebellar, striatal and hippocampal slices: a manifestation of spreading depression?

Spreading depression, which can be evoked by a variety of stimuli both in vitro and in vivo, is associated with profound changes in extracellular ion concentrations and enhanced release of neurotransmitter amino acids. We have observed a transient spontaneous release of amino acids in slice preparations obtained from rat cerebellum, striatum and hippocampus; this phenomenon has similar properties to stimulus-evoked spreading depression. Aspartate, glutamate, glutamine, serine, glycine and γ-aminobutyric acid (GABA) release were potentiated during these episodes in all three brain regions, with a variable effect upon taurine release. When compared to glutamate release, a consistently high release of aspartate, glycine and serine was observed. Amino acid release, evoked by whole slice depolarization using veratridine (10–25 μM) or elevated potassium (35–60 mM) consistently enhanced glutamate release, and to a lesser extent aspartate release, but had negligible effect upon the other amino acids. Thus, the release profiles for spontaneous and depolarization-evoked release are markedly different. We suggest that the spontaneous release observed in brain slices represents a spreading depression-like phenomenon; the putative roles of the amino acids are discussed.

Profound systemic hypothermia inhibits the release of neurotransmitter amino acids in spinal cord ischemia

Profound hypothermia induced with cardiopulmonary bypass has a protective effect on spinal cord function during operations on the thoracoabdominal aorta. The mechanism of this protection remains unknown. It has been proposed that the release of excitatory amino acids in the extracellular space plays a causal role in irreversible neuronal damage. We investigated the changes in extracellular neurotransmitter amino acid concentrations with the use of in vivo microdialysis in a swine model of spinal cord ischemia. All animals underwent left thoracotomy and right atrium-femoral artery cardiopulmonary bypass with additional aortic arch perfusion. Lumbar laminectomies were then done and microdialysis probes were inserted stereotactically in the anterior horn of the second and fourth segments of the lumbar spinal cord. The probes were perfused with artificial cerebrospinal fluid at a rate of 2 microliters/min and 15-minute samples were assayed by high-performance liquid chromatography. Group 1 animals (n = 6) underwent aortic clamping distal to the left subclavian artery and proximal to the renal arteries for 60 minutes at normothermia (37 degrees C) and group 2 animals (n = 5) were cooled to a rectal temperature of 20 degrees C before application of aortic clamps, maintained at this level during cardiopulmonary bypass until the aorta was unclamped, and then slowly rewarmed to 37 degrees C. Seven amino acids were studied, including two excitatory neurotransmitters (glutamate and aspartate) and five putative inhibitory neurotransmitters (glycine, gamma-aminobutyric acid, serine, adenosine, and taurine). Glutamate exhibited a threefold increase in extracellular concentration during normothermic ischemia compared with baseline values and remained elevated until 60 minutes after reperfusion. The increase in aspartate concentration was not significant. The extracellular concentrations of glycine and gamma-aminobutyric acid also increased significantly during ischemia and reperfusion. Hypothermia uniformly prevented the release of amino acids in the extracellular space. Glutamate levels remained significantly decreased even after rewarming to normothermia whereas glycine levels returned to baseline values. These results are consistent with a role for excitatory amino acids in the production of ischemic spinal cord injury and suggest that the mechanism of hypothermic protection may be related to decreased release of these amino acids in the ischemic spinal cord.

A possible role for phospholipases in the release of neurotransmitter amino acids from ischemic rat cerebral cortex

The involvement of phospholipases in ischemia-evoked release of aspartate, glutamate, glycine, and GABA from the cerebral cortex was studied in a four vessel occlusion rat model of cerebral ischemia/reperfusion. In comparison with the control group, the phospholipase A 2 inhibitor mepacrine significantly decreased the ischemia-evoked efflux of transmitter amino acids into cortical superfusates. Direct application of phospholipases A 2 or C to the cerebral cortex of non-ischemic animals resulted in a significant increase in amino acid levels. These results suggest that neurotransmitter release following cerebral ischemia may involve phospholipase induced plasma membrane disruption.

Glucagon-like peptide-1(7–36)amide induces the release of aspartic acid and glutamine by the ventromedial hypothalamus of the conscious rat

Glucagon-like peptide-1(7–36)amide (GLP-1(7–36)amide) and its own receptor have been found in the hypothalamus and brain stem of the rat In an attempt to gain further insight into the role of this peptide in brain functioning we investigated the effects of GLP-1(7–36)amide on the release of excitatory amino acid neurotransmitters by the ventromedial hypothalamus using an experimental microdialysis approach. GLP-1(7–36)amide produced an immediate increase in the extracellular concentrations of aspartic acid and glutamine, p < 0.01 and p < 0.05, respectively. By contrast, extracellular concentrations of glutamic acid, alanine, threonine, and tyrosine were unaffected. The results of this study show a stimulatory effect of GLP-1(7–36)amide on the release of aspartic acid and glutamine by the ventromedial hypothalamus of the rat.

The effect of electrical head-only stun duration on electroencephalographic-measured seizure and brain amino acid neurotransmitter release

Effective electrical head-only stunning produces a seizure-like state followed by a period of analgesia seen in animals allowed to recover. Passing of a 1·0 A current (50 Hz, 500 V) for less than 0·2 s, through the head of a sheep does not produce a seizure-like state as evidenced by recorded electroencephalogram. Corresponding to this lack of seizure-like state, the release of the neurotransmitters glutamate and aspartate, in the brain, occurs to levels associated with arousal rather than seizure. At a duration of 0·2 s, the same stun parameters as above produce a seizure-like state and the release of glutamate and aspartate rises dramatically. The length of the seizure-like state, and the levels of release of glutamate, aspartate and a third neurotransmitter gamma amino-4-butyric acid (GABA), increased with stun duration until 4·0 s duration, where a peak in these parameters was seen. Stun durations of 2·0, 4·0, 8·0 and 12·0 s all produce similar effects. At a duration of 20 s, however, the length of the seizure-like state and the release of neurotransmitters is less than at shorter stun durations. For welfare purposes a head-only electrical stun, of 1·0 A, at a duration as low as 0·2 s produces unconsciousness and analgesia to subsequent slaughter procedures. However, maximum welfare benefits appear attainable at durations between 2·0 and 20·0 s.

Preoptic rather than mediobasal hypothalamic amino acid neurotransmitter release regulates GnRH secretion during the estrogen-induced LH surge in the ovariectomized rat.

Inhibitory and excitatory amino acid neurotransmitters have been suggested to participate in the feedback actions of estradiol (E2) on LH secretion. In the rat estrogen-receptive neurons have been demonstrated in the preoptic/anterior hypothalamic area (POA) and mediobasal hypothalamus/median eminence (MBH) and many of these neurons utilize gamma-aminobutyric acid (GABA) as neurotransmitter. The actions of excitatory amino acids (EAA) differ in ovariectomized (ovx) and ovx E2-substituted rats indicating that EAAs also participate in the positive feedback action of E2 on LH release. However, little information is available as to whether in vivo these transmitters exert their effects in the POA, where most of the GnRH perikarya are located, or in the MBH, i.e. at the nerve terminals. Therefore we conducted push pull cannula perfusions to compare the release rates of GABA, aspartate (ASP) and glutamate (GLU) in the MBH and POA. A subcutaneous implant of a silastic tube containing E2 resulted in LH surges in the afternoon of all treated animals. Prior to and during this LH surge the MBH release rates of neither GABA nor ASP nor GLU were significantly altered. In contrast, a conspicuous drop in preoptic GABA release occurred prior to and during the time of estrogen-induced LH surges and this was accompanied by enhanced preoptic secretion of ASP and GLU. In conclusion, we present the first data about amino acid release in the MBH during the E2-induced LH surge. Since only in the POA the LH surge is associated with changes in amino acid release, it appears that both inhibitory and excitatory amino acids act at the level of the GnRH cell bodies and/or dendrites and not on GnRH nerve terminals to mediate the feedback mechanism of E2 on LH release.

LTP in the dentate gyrus is associated with a persistent NMDA receptor-dependent enhancement of synaptosomal glutamate release

LTP was induced unilaterally in the dentate gyrus of the anaesthetized rat by tetanic stimulation of the perforant path. Two hours after tetanization synaptosomes were prepared from control and potentiated sides, and the depolarization-induced release of endogenous amino acid neurotransmitters was compared in the two groups, using 4-aminopyridine, potassium or veratridine as depolarizing agents. Glutamate release was greater in synaptosomes from potentiated than control tissue with all three depolarizing agents, but the highest release from control synaptosomes and the most significant enhancement of release from potentiated synaptosomes was seen with veratridine. The tetanus-induced enhancement of veratridine-induced glutamate release was prevented by pretreatment with MK-801. The results indicate that the induction of LTP in the dentate gyrus is accompanied by a persistent enhancement in the ability of synaptosomes to release glutamate in response to a depolarizing stimulus.

The effects of sigma ligands on the release of glutamate from rat striatal slices

This study investigated the effects of sigma receptor ligands on the release of endogenous amino acid neurotransmitters from rat striatal slices. The effect of haloperidol on release in slices prepared from 6-hydroxydopamine lesioned animals was also tested. Haloperidol, the (+/-) reduced metabolite of haloperidol, rimcazole and ifenprodil specifically reduced potassium-stimulated release of glutamate with IC50 values between 20-60 microM. The release of aspartate, gamma-aminobutyric acid (GABA) and glycine was not affected. Haloperidol also reduced glutamate release from slices prepared from lesioned animals. The neuroleptic drug alpha-flupenthixol and the putative sigma receptor ligand R(+)3-(3-hydroxyphenyl)-N-(n-propyl) piperidine (3-PPP) had no effect on release. These effects of the sigma ligands show that the inhibition of glutamate release is specific to this amino acid and also that it is not due to dopamine receptor blockade as those ligands which have low affinity for dopamine receptors were also effective in reducing release. A presynaptic location for sigma receptor sites, possibly associated with ion channels, could account for the effects of these ligands on transmitter release.

Free radicals enhance basal release of D-[3H]aspartate from cerebral cortical synaptosomes.

Excessive generation of free radicals has been implicated in several pathological conditions. We demonstrated previously that peroxide-generated free radicals decrease calcium-dependent high K(+)-evoked L[3H]-glutamate release from synaptosomes while increasing calcium-independent basal release. The present study evaluates the nonvesicular release of excitatory amino acid neurotransmitters, using D-[3H]aspartate as an exogenous label of the cytoplasmic pool of L-glutamate and L-aspartate. Isolated presynaptic nerve terminals from the guinea pig cerebral cortex were used to examine the actions and interactions of peroxide, iron, and desferrioxamine. Pretreatment with peroxide, iron alone, or peroxide with iron significantly increased the calcium-independent basal release of D-[3H]aspartate. Pretreatment with desferrioxamine had little effect on its own but significantly limited the enhancement by peroxide. High K(+)-evoked release in the presence of Ca2+ was enhanced by peroxide but not by iron. These data suggest that peroxide increases nonvesicular basal release of excitatory amino acids through Fenton-generated hydroxyl radicals. This release could cause accumulation of extracellular excitatory amino acids and contribute to the excitotoxicity associated with some pathologies.

Effects of apomorphine and L-methionine sulphoximine on the release of excitatory amino acid neurotransmitters and glutamine in the striatum of the conscious rat.

The effects of apomorphine, a D1-, D2-dopamine receptor agonist, on the extracellular concentrations of excitatory amino acids, glutamic and aspartic acids, and on that of their precursor, glutamine, were investigated using an intracerebral perfusion system. Apomorphine produced a concentration-related rise in glutamic acid concentration in cerebral perfusates (P < 0.01) whereas only the highest concentration of apomorphine (3 x 10(-3) micrograms/microliters) increased the concentration of aspartic acid (P < 0.05). These effects were seen in the sample taken at the same time as the apomorphine injection. The rise in glutamine concentration (P < 0.01) produced by apomorphine continued for 10 min beyond perfusion with apomorphine. These effects were attenuated by previous injections of D1-, D2-dopamine receptor blocker. To investigate further the release of glutamine, the glutamine synthetase inhibitor L-methionine sulphoximine (MSO) was injected intracerebrally before apomorphine perfusion. After MSO pre-injection, the extracellular concentration of glutamine decreased (P < 0.01) to near zero concentrations. In MSO-treated animals, apomorphine did not induce the release of glutamic acid, aspartic acid or glutamine. These results indicate a role for dopamine in the release of excitatory amino acids and glutamine in the neostriatum of the rat. A possible volumetric interaction between dopamine and glutamic acid as well as the hypothesis of a striato-pallido-thalamo-cortico-striatal feedback loop are discussed.

The effect of neuropeptides on the release of neurotransmitter amino acids from rat striatum

Neuropeptide Y (NPY), peptide YY (PYY), and galanin are found throughout the central nervous system with appreciable levels occurring in the striatum. In this study we have investigated the effects of these peptides on the potassium-stimulated release of endogenous neurotransmitter amino acids from slices of rat striatum. The release of glutamate was significantly reduced by nanomolar concentrations of each peptide, whilst the release of aspartate, γ-aminobutyric acid (GABA) and glycine was not affected. The reduction in release due to galanin (200 nM) was inhibited by glibenclamide (5 μM). These results support the view that NPY, PYY and galanin modulate neurotransmitter release possibly by a presynaptic action. The results with glibenclamide suggest that the action of galanin is mediated through and ATP-sensitive potassium channel.

Characterization of glutamate, aspartate, and GABA release from ischemic rat cerebral cortex

The purpose of this study was to evaluate potential mechanisms of ischemia-evoked amino acid transmitter release. Changes in extracellular levels of transmitter amino acids and lactic acid dehydrogenase (LDH) in rat cerebral cortex during and following four-vessel occlusion elicited global cerebral ischemia were examined using a cortical cup technique. Ischemia-evoked release of glutamate, aspartate and γ-amino-butyric acid (GABA) was compared in control vs. drug-treated animals. Tetrodotoxin and antagonists of glutamate receptors (DNQX, MK-801, and AP-3) depressed the initial rate of increase in extracellular glutamate and aspartate without altering the total amount of these amino acids collected in the conical superfusates. Cobalt, a calcium channel antagonist, failed to alter efflux. Acidic amino acid transport inhibitors (dihydrokainate, L-trans-PDC) depressed the rate of onset of glutamate and aspartate release and dihydrokainate depressed total release by 44%. PD 81723, an allosteric enhancer at the A, adenosine receptor, depressed glutamate efflux, as did L-NAME, an inhibitor of nitric oxide synthase. Extracellular increases in GABA levels were depressed by tetrodotoxin and L-trans-PDC. The GABA transport inhibitor, nipecotic acid, increased the initial rate of onset of GABA release. Increases in LDH levels in the extracellular fluid became apparent during the period of ischemia and continued to increase during the subsequent 90 min of reperfusion. These results suggest that ischemia evokes a release of neurotransmitter amino acids that is only partially dependent upon Ca 2+ influx activation or the reversal of amino acid transporters. Nonselective mechanisms, resulting from the disruption of plasma membrane integrity, may contribute significantly to the total ischemia-evoked release of excitatory amino acids.

Effects of amphetamine on the release of excitatory amino acid neurotransmitters in the basal ganglia of the conscious rat.

The effects of systemic injections of amphetamine sulfate on the release of aspartic acid, glutamic acid, and glutamine were studied using a push-pull perfusion system in the conscious rat. Amphetamine produced a dose-related increase of the extracellular levels of aspartic acid and glutamic acid. The mean time effect of amphetamine was 40 min, followed by a recovery to baseline levels. The mean percentage increase in amino acids released by the highest dose of amphetamine (5 mg/kg) was as follows: Asp, 334.6%; Glu, 224.5%; and Gln, 317.6%. All these effects were blocked by the dopamine D1-D2 receptor blocker haloperidol. It is suggested that dopamine, released by amphetamine, induces the release of amino acid neurotransmitters in the neostriatum. In addition, it is proposed that dopamine could mediate the neurotoxic effects produced by amphetamines through their secondary action on the release of excitatory amino acids.

Adenosine modulation of amino acid release in rat hippocampus during ischemia and veratridine depolarization

This study was undertaken to determine whether endogenous adenosine modulates ‘in vivo’ neurotransmitter amino acid release via its presynaptic receptors. Two conditions were compared: neuronal depolarization by local infusion of veratridine (600 μM), and transient global ischemia by four-vessel occlusion. Both stimuli were applied for 20 min. Extracellular amino acid (glutamate, taurine/GABA, glycine) variations in concentration were determined in the rat hippocampus by microdialysis and HPLC. Modulation of adenosine receptor activity was objectified by continuous local infusion of an adenosine agonist ( R-phenylisopropyladenosine R-PIA) or an antagonist (theophylline), starting one hour before stimulation of amino acid release. R-PIA (100 μM) significantly decreased the glutamate release (50%) evoked by veratridine, whereas it did not significantly modify the ischemia-induced glutamate release. In contrast, theophylline did not significantly affect veratridine-induced glutamate release, but it significantly potentiated glutamate efflux (400%) under ischemic conditions. Neither treatment altered the release of the other amino acids. These data suggest that endogenous adenosine appearing in the extracellular space during veratridine-induced depolarization cannot control glutamate release. In contrast, ischemia-induced glutamate release was strongly inhibited by the concomitant increase in extracellular adenosine.

Aging, prefrontal cortex, and amino acid neurotransmitters: Differential effects produced by electrical stimulation

The purpose of the present study was to investigate the effects of electrical stimulation on the in vivo release of amino acid neurotransmitters in the prefrontal cortex of young (3–4 months) and aged (27–30 months) rats. In vivo push-pull perfusions were performed in the conscious animal. The levels of aspartate, glutamate, and glycine were analyzed by HPLC-fluorimetric detection. Electrical stimulation of the medial prefrontal cortex at the intensity of 0.3 mA produced a significant increase in the levels of aspartate, glutamate, and glycine in young but not aged rats. However, at the intensity of 0.6 mA, a significant delayed increase of these same amino acids was produced in the aged rats. These results support the existence of a differential effect of electrical stimulation on amino acid neurotransmitters in the prefrontal cortex of young and aged rats.

Amino acid levels and gamma-aminobutyric acidA receptors in rat neostriatum, cortex, and thalamus after neonatal 6-hydroxydopamine lesion.

The amino acid gamma-aminobutyric acid (GABA) is the main inhibitory neurotransmitter in brain, and GABAergic neurons have been proposed to play a major role in basal ganglia physiology. In the neostriatum (caudate putamen), medium-sized aspiny interneurons, as well as neostriatal output neurons that project to several brain regions, use GABA as their neurotransmitter. Dopamine fibers arising from the substantia nigra represent a major input to the neostriatum where, besides their classic neurotransmitter role, they are seemingly involved in the regulation of amino acid neurotransmitter release. To further characterize the nature of some of the amino acid/dopamine interactions, selective dopaminergic deafferentations were produced in neonatal rats (3 days postnatal) by intraventricular administration of the neurotoxin 6-hydroxydopamine (6-OHDA); the noradrenergic neurons were protected by prior administration of desmethylimipramine. After a 3-month survival, levels of catecholamines, indoleamines, and amino acids were determined in cingulate cortex, thalamus, and neostriatum. In addition, GABAA receptors were measured in membrane preparations from these three regions, using the specific agonist [3H]muscimol. In the 6-hydroxydopamine-lesioned rats, levels of dopamine and its metabolites homovanillic acid, 3,4-dihydroxyphenylacetic acid, and 3-methoxytyramine were decreased, as expected, in cortex and neostriatum, but remained unmodified in thalamus. In all three regions, serotonin content was increased; its metabolite, 5-hydroxyindole-3-acetic acid, was also elevated, but only in cortex and neostriatum. The levels of GABA were increased in neostriatum and thalamus, but remained unmodified in cortex. Glycine was increased in all three regions examined. There were also increases of phosphatidylethanolamine and serine in thalamus, and of aspartic acid and alanine in neostriatum. The density of GABAA binding sites was increased in neostriatum, but remained unchanged in cortex and thalamus. The changes in amino acid levels and [3H]muscimol binding sites induced by a neonatal 6-hydroxydopamine treatment differ from those found after similar lesions in adult animals, possibly because of the plastic and synaptic rearrangements that can still occur during early postnatal development. The present results also demonstrate that adaptations occur in response to a dopaminergic deafferentation at an early age and that these exhibit a regional specificity.

Preoptic release of amino acid neurotransmitters evaluated in peripubertal and young adult female rats by push-pull perfusion.

In order to evaluate the participation of amino acid neurotransmitters (AANT) in pubertal development, we measured the preoptic release rates of glutamate (Glu), aspartate (Asp), homocysteic acid (HCA), gamma-aminobutyric acid (GABA), glycine (Gly), and taurine (Tau) in female rats aged 26, 35 and 42 days. Push-pull cannulae were implanted in the preoptic area (POA) after reinforcing the os calvarium by cementing a slightly curved 2-mm thick polyethylene plaque. Four days later, 120-min perfusion experiments (0-90 min: artificial CSF; 90-120 min: CSF containing 50 mM KCl) were performed. Amino acid concentrations were measured in 15-min fractions after derivatization with phenylisothiocyanate, using reverse-phase high-performance liquid chromatography and UV detection. Rats aged 42 days showed significantly (p < 0.01) higher preoptic Asp release rates, as compared to 26-day-old rats; Glu output increased significantly (p < 0.05) in animals aged 35 and 42 days, as compared to 26-day-old rats. No significant changes in HCA release were observed. Preoptic release rates of GABA, Gly and Tau were significantly higher in 26-day-old animals as compared to 35-day-old (GABA, p < 0.05) and 35- and 42-day-old rats (Gly, Tau, p < 0.01). Basal release of Asp, Glu, HCA, Tau and GABA, but not of Gly, was significantly stimulated (p < 0.05) by KCl in all age groups. It is suggested that in peripubertal female rats, increased preoptic Asp and Glu release rates and decreased GABA and Tau output might enhance luteinizing hormone-releasing hormone secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Dopamine receptor antagonist blocks the release of glycine, GABA, and taurine produced by amphetamine

The effects of systemic injections of amphetamine sulfate on the extracellular levels of glycine, GABA, and taurine in the neostriatum of awake rats were studied using a push-pull perfusion system. Amphetamine produced a dose-related increase in glycine levels. Amphetamine also produced an enhancement on GABA and taurine levels, although these increases did not follow a dose-related curve. The percentage increase of amino acids produced by the highest dose of amphetamine (5 mg/kg) at the peak effect was: GLY 235.9%; GABA 218%, and TAU 177%. All these effects were blocked by the D1–D2 dopamine receptor antagonist, haloperidol. It is suggested that dopamine, released by amphetamine, induces the release of inhibitory amino acid neurotransmitters in the neostriatum. These results are consistent with the hypothesis of dopamine playing a role of an amplifier of the activity of different neurochemical circuits. The results are also in accord with the idea that dopamine could mediate the neurotoxic effects produced by amphetamines through an interplay between excitatory and inhibitory amino acids.