Changes in the expression of glial glutamate transporters in the rat brain accompanied with morphine dependence and naloxone-precipitated withdrawal

The expression of mRNAs for the glial glutamate transporters, GLT-1 and GLAST, in the rat brain accompanied with morphine dependence and naloxone-precipitated withdrawal was investigated by Northern blot analysis. The expression of GLT-1 mRNA was significantly decreased in the striatum and thalamus of morphine-dependent rats, and significantly increased in the striatum 2 h after the naloxone-precipitated withdrawal, compared with that of naive rats. On the other hand, there were no significant changes in GLAST mRNA level in any brain region. These results suggest the involvement of GLT-1 in the development of morphine dependence and the expression of morphine withdrawal.     

大量饮酒及乙醇戒断后对大鼠脑内维生素C水平的影响

目的研究大量饮酒4 d及乙醇戒断后大鼠脑脊液中及各重要脑区内维生素C(VC)水平及钠依赖性VC转运体2(SVCT2)蛋白水平的变化。方法30只健康Wistar大鼠随机分为5组,对照组(A组)、大量饮酒4 d(B组)、大量饮酒4 d后戒断1 d(C组)、大量饮酒4 d后戒断2 d(D组)、大量饮酒4 d后戒断7 d(E组),每组6只。B、C、D、E组大鼠大量灌胃给予乙醇4 d,乙醇浓度为25%W/V,每8小时灌胃1次,连续4 d;A组给予等体积蒸馏水。采用Y迷宫实验评价大鼠的空间工作记忆能力;以高效液相-电化学法(HPLC-ECD)检测大鼠脑脊液及前额叶皮质、顶叶皮质、颞叶皮质、海马脑区内细胞内VC含量,免疫印迹法检测大鼠各脑区内SVCT2蛋白水平的变化。结果(1)Y迷宫实验:C、D组大鼠自发交替反应率分别为(24.64±15.11)%和(41.48±13.01)%,均显著低于A组[(75.47±8.61)%];E组大鼠的自发交替反应率恢复接近正常[(61.64±11.51)%],与A组差异无统计学意义(P>0.05)。(2)脑脊液内VC含量:B组大鼠脑脊液内VC含量为(204.54±25.51)μmol/L,显著高于A组[(145.57±18.98)μmol/L],而C、D组大鼠脑脊液内VC水平[分别为(90.24±15.45)、(86.93±14.53)μmol/L]明显低于A、B组,差异均有统计学意义(均P<0.001);E组大鼠脑脊液中VC含量[(135.80±17.16)μmol/L]与A组间差异无统计学意义(P>0.05)。(3)各脑区内VC水平:B组前额叶皮质、顶叶皮质、海马脑区脑组织匀浆(细胞内)VC水平变化趋势相同[分别为(1.18±0.13)、(1.14±0.12)、(1.20±0.20)μmol/g],均显著低于A组[分别为(1.64±0.11)、(1.62±0.13)、(2.06±0.27)μmol/g];C组在乙醇戒断后,前额叶皮质、顶叶皮质、海马脑区细胞内VC水平[分别为(1.20±0.29)、(1.05±0.06)、(1.21±0.15)μmol/g]有所恢复,但仍明显低于A组(均P<0.01);C组大鼠颞叶皮质细胞内VC水平[(1.37±0.04)μmol/g]显著高于顶叶皮质(P<0.05);D组各脑区细胞内VC水平均有回升趋势,前额叶皮质细胞内VC水平显著高于顶叶皮质[(1.63±0.24)μmol/g比(1.26±0.16)μmol/g,P<0.05];E组前额叶皮质、顶叶皮质、颞叶皮质、海马区细胞内VC水平[分别为(1.72±0.19)、(1.43±0.22)、(1.67±0.19)、(1.86±0.22)μmol/g]均较B组显著升高(均P<0.01),逐渐恢复至正常水平,海马区VC水平显著高于顶叶皮质(P<0.05)。(4)Western blot结果显示,与A组比较,B、C、D组大鼠前额叶皮质、顶叶皮质及C、D、E组大鼠海马区的SVCT2蛋白均显著升高(均P<0.05)。结论大量饮酒4 d可致大鼠脑损伤,乙醇干预后大鼠脑脊液中及各脑区脑组织匀浆中VC水平呈现负相关的变化过程,此过程中SVCT2蛋白表达上调,这将有助于脑脊液内的VC被转运至各脑区神经元内发挥抗氧化作用。     

Effects of agmatine on ethanol withdrawal syndrome in rats

Effects of agmatine, which is an endogenous polyamine metabolite formed by decarboxylation of L-arginine, have been investigated on the ethanol withdrawal syndrome in rats. Adult male Wistar rats were used in the study. Ethanol (7.2% v/v) was given to the rats by a liquid diet for 21 days. Agmatine (20, 40, 80 and 160 mg/kg) and saline were injected to rats intraperitoneally 30 min before ethanol withdrawal testing. After 30th min, 2nd and 6th h of ethanol withdrawal, rats were observed for 5 min, and withdrawal signs which included locomotor hyperactivity, agitation, stereotyped behavior, wet dog shakes and tremor were recorded or rated. A second series of injections was given at 6 h after the first one, and subjects were then tested for audiogenic seizures. Agmatine caused dose-dependent and significant inhibitory effects on stereotyped behaviors, wet dog shakes and tremors during the observation period. It did not cause any significant change in motor coordination of naive (not ethanol-dependent) rats. Our results suggest that agmatine attenuates withdrawal syndrome in ethanol-dependent rats; thus, this drug may be beneficial in the treatment of ethanol dependence.     

Effects of fluoxetine on ethanol withdrawal syndrome in rats

The present study was designed to investigate the effects of fluoxetine, a selective serotonin reuptake inhibitor, on ethanol withdrawal syndrome in rats. Adult male Wistar rats (218-255 g) were subjects. Ethanol (7.2%, v/v) was given to rats by a liquid diet for 21 days. Control rats were pair fed an isocaloric liquid diet containing sucrose as a caloric substitute to ethanol. Fluoxetine (2.5, 5 and 10 mg/kg) and saline were injected to rats intraperitoneally just before ethanol withdrawal. After 2nd, 4th and 6th hour of ethanol withdrawal, rats were observed for 5 min, and withdrawal signs that included locomotor hyperactivity, agitation, stereotyped behavior, wet dog shakes and tremor were recorded or rated. A second series of injections was given at 6 h after the first one, and subjects were then tested for audiogenic seizures. Fluoxetine produced some dose-dependent and significant inhibitory effects on all the signs of ethanol withdrawal during ethanol withdrawal period. Our results suggest that acute fluoxetine treatment has some beneficial effects on ethanol withdrawal in rats. Thus, this drug may be useful for treatment of ethanol withdrawal syndrome.     

Effects of escitalopram on ethanol withdrawal syndrome in rats

The present study was designed to investigate the effects of escitalopram, a selective serotonin reuptake inhibitor, on ethanol withdrawal syndrome in rats. Adult male Wistar rats (266-278 g) were subjects. Ethanol (7.2%, v/v) was given to rats by a liquid diet for 21 days. Control rats were pair fed with an isocaloric liquid diet containing sucrose as a caloric substitute to ethanol. Escitalopram (2.5, 5 and 10 mg/kg) and saline were injected to rats intraperitoneally just before ethanol withdrawal. After the second and sixth hours of ethanol withdrawal, rats were observed for 5 min, and withdrawal signs that included locomotor hyperactivity, agitation, stereotyped behavior, wet dog shakes, tremors and audiogenic seizures were recorded or rated. A second series of injections was given 30 min before sixth hour of withdrawal test. Effects of escitalopram on the locomotor activities of the na?ve (no ethanol-dependent) rats were also evaluated. Escitalopram (5 mg/kg) reduced the increased stereotyped behaviors at the sixth hour of ethanol withdrawal. It inhibited tremors at the second hour of ethanol withdrawal at doses of 5 and 10 mg/kg. Escitalopram (2.5 and 5 mg/kg) also produced some significant attenuations in the incidence of wet dog shakes at the second and sixth hours of the observation period. It was found ineffective on locomotor hyperactivity, agitation and audiogenic seizures. Escitalopram (2.5 and 5 mg/kg) did not cause any significant effect on locomotor activities of the na?ve rats. Our results suggest that acute escitalopram treatment has some limited beneficial effects on ethanol withdrawal syndrome in rats.     

Effects of in utero ethanol exposure on the developing dopaminergic system in rats

Previous studies from this and other laboratories suggest that dopamine is decreased in selected brain regions of postnatal rats exposed to ethanol in utero. The present study expands previous work by examining the effects of in utero ethanol exposure on dopamine D1 and D2 binding sites and dopamine uptake in postnatal rats. In addition, dopamine content in the brain stem and frontal cortex of fetal and neonatal rats was examined. The experimental results indicate that in utero ethanol exposure markedly affects the postnatal development of the dopaminergic system in the striatum and frontal cortex. We observed a marked, transient deficiency of striatal dopamine (greater than 40% decrease at 19 days) and dopamine uptake sites (approximately 25% decrease in Vmax at 35 days). The Bmax for striatal dopamine D1 binding sites was decreased by greater than 20% at both 19 and 35 days. Cortical D1 sites were markedly decreased at 19 days (greater than 40%). In contrast, the number of striatal D2 receptors was unaffected by in utero ethanol exposure at both ages. Analysis of tissue from neonatal rats demonstrated a marked dopamine deficiency in ethanol-exposed rats on postnatal day 5. In light of the proposed morphogenic actions of dopamine early in development, it is possible that the early dopamine deficiency contributes to the abnormal postnatal development of the dopaminergic system.     

Effects of risperidone, quetiapine and ziprasidone on ethanol withdrawal syndrome in rats

Comorbid substance use in schizophrenic patients is common, and substance dependence is a predictive factor for psychosis. The present study was designed to investigate the effects of risperidone, quetiapine and ziprasidone, atypical antipsychotic drugs, on ethanol withdrawal syndrome (EWS) in rats. Adult male Wistar rats were used in the study. Ethanol (7.2%, v/v) was given to rats via a liquid diet for 21 days. An isocaloric liquid diet without ethanol was given to control rats. Risperidone (1 and 2 mg/kg), quetiapine (8 and 16 mg/kg), ziprasidone (0.5 and 1 mg/kg) and vehicle were injected into rats intraperitoneally at 1.5 and 5.5 h of ethanol withdrawal. At the 2nd, 4th and 6th hours of ethanol withdrawal, rats were observed for 5 min, and withdrawal signs that included locomotor hyperactivity, stereotyped behaviors, abnormal gait and posture, tail stiffness and agitation were recorded or rated. Following the observations at the 6th hour, the rats were tested for audiogenic seizures. All three drugs had some significant inhibitory effects on EWS-induced behavioral signs beginning at the 2nd hour of withdrawal. The drugs also significantly reduced the incidence of audiogenic seizures. Overall, risperidone and quetiapine seemed to be more effective than ziprasidone in ameliorating the withdrawal signs. Doses of the drugs used in the present study did not produce any significant changes in locomotor activities of naïve rats. Our results suggest that risperidone, quetiapine and ziprasidone had beneficial effects on EWS in rats. Thus, these drugs may be helpful for controlling withdrawal signs in ethanol-dependent patients.     

Downregulation of glial glutamate transporters after dopamine denervation in the striatum of 6-hydroxydopamine-lesioned rats

Overactivity of glutamatergic neurotransmission in the basal ganglia is known to be closely related to the onset and pathogenesis of Parkinson's disease. Glutamate homeostasis around glutamatergic synapses is tightly regulated by two groups of glutamate transporters: glial glutamate transporters GLT1 (EAAT2) and GLAST (EAAT1), and neuronal glutamate transporter EAAC1. In order to investigate the changes of glutamate transporters after the onset of Parkinson's disease, unilateral 6-hydroxydopamine-lesioned rat, an animal model of Parkinson's disease, was employed. By immunofluorescence and Western blot analyses, GLT1 and GLAST proteins were significantly reduced in the striatum with lesion. No change in GLT1 and GLAST protein was found in the substantia nigra. The reduction of GLT1 protein in the striatum was more prominent than that of GLAST protein (approximately 40% vs. 20%). In addition, EAAC1 protein was found to be increased in the substantia nigra pars reticulata of the lesioned rats but not in the striatum. The present results indicate that reductions of GLT1 and GLAST may impair glutamate homeostasis around glutamatergic synapses in the striatum and contribute to over-spills of glutamate in the system. An increase in the EAAC1 level in the substantia nigra pars reticulata may increase GABA synthesis and enhance GABAergic neurotransmission. These results indicate that there are differential and distinct modulations of glutamate transporters after dopamine denervation in the 6-hydroxydopamine-lesioned rat.     

Transient expression of the glial glutamate transporters GLAST and GLT in hippocampal neurons in primary culture

The extracellular glutamate concentration is kept low by glutamate transporters in the plasma membranes. Here we have studied the expression of the glutamate transporters GLAST, GLT and EAAC during the in vitro development of embryonic hippocampal neurons grown in a defined (serum free) medium. Immunochemistry studies showed that both the GLAST and GLT proteins are expressed in a subpopulation of neurons at the early, but not at the later stages of the cultures. Glial cells expressing the GLAST and GLT proteins were found at all stages. EAAC was only detected in neurons. This is one of the first evidence of a neuronal ability to express GLAST.     

Changes in expression of neuronal and glial glutamate transporters in rat hippocampus following kainate-induced seizure activity

The expression of excitatory amino acid transporters (EAATs) in rat hippocampus was studied following kainic acid-induced seizure activity in vivo and in hippocampal slice cultures. Protein and mRNA levels of the glial (EAAT2) and neuronal (EAAT3) transporters were determined with affinity-purified antibodies and oligonucleotide probes, respectively. Kainate treatment decreased EAAT3 immunoreactivity in stratum lacunosum moleculare within 4 h of seizure onset. Upon pyramidal cell death (5 days after kainate treatment), EAAT3 immunoreactivity in stratum pyramidale of CA1 and in stratum lacunosum moleculare was almost completely eliminated. The rapid effect of kainate on EAAT3 expression was confirmed by in situ hybridization; EAAT3 mRNA levels were decreased in CA1 and CA3 regions within 4-8 h of seizure onset. Kainate treatment had an opposite effect on levels and expression of EAAT2. Developmental studies indicated that the rapid regulation of transporter expression was not observed in rats younger than 21 days, an observation congruent with previous reports regarding the resistance of young rats to kainate. In hippocampal organotypic cultures, which lack a major excitatory input from the entorhinal cortex, kainate produced a slow decrease in [3H]d-aspartate uptake. This study indicates that an early effect of kainate treatment consists of down-regulation of the neuronal transporter EAAT3 in restricted hippocampal regions, together with a modest increase in the expression of the glial transporter EAAT2. Differential regulation of neuronal and glial glutamate transporters may thus play a role in kainate-induced seizure, neurotoxicity and neuronal plasticity.     

Effects of postnatal ethanol exposure on glial cell development in rat optic nerve

This study morphologically evaluated the effects of limited postnatal alcohol exposure on the development of glial cells in the rat optic nerve. Rat pups were artificially reared on Days 5-18 with a supplemented milk diet fed via a chronic gastrostomy tube. Experimental animals received 4% ethanol in their diet on Days 5-9, otherwise the experimental and control animals received identical diets. Optic nerve tissues were prepared for electron microscopy on Days 10, 16, 22, 29, and 90. There were fewer glial cells per cross section and the cross-sectional areas of optic nerves were smaller on Days 10 and 16 in the ethanol-exposed animals. The alcohol caused a delay in the maturation of oligodendroglial cells at 10 days as evidenced by decreases in the total number of oligodendroglia present and by a delay in the appearance of immature cells within the oligodendroglial lineage. All of these effects were compensated for at later ages. There was no evidence of alcohol-induced degeneration of glial cells or their organelles. Thus, postnatal alcohol exposure causes a delay in oligodendrocyte maturation but appears to have no long-term effects on the glial cell population of rat optic nerve.     

Expression of high-affinity neuronal and glial glutamate transporters in the rat optic nerve

Recent studies have revealed that a dynamic axon-glial signaling occurs in the rat optic nerve, which is devoid of synapses. This interaction is postulated to be mediated by non-vesicular release of glutamate via a reversal of high-affinity glutamate transporters. Here we examined the expression of glial glutamate transporters (GLAST and GLT-1) and a neuronal transporter (EAAC1) in the rat optic nerve. RT-PCR analysis revealed the presence of mRNAs for GLT-1 and GLAST, but not EAAC1. RNase protection assays showed that of the two glial transporters, mRNA for GLAST was expressed at much higher level than was GLT-1. A similar expression pattern was found in primary astrocyte culture cells. GLAST mRNA level in the optic nerve was comparable to that in the cerebellum. Developmentally, GLAST mRNA level was highest at P2 and dropped slightly by adulthood. Nerve transection resulted in little or no change in mRNA levels for GLAST and GLT-1 assayed at 4 to 14 days post-transection, but GLAST mRNA level was decreased at 64 days. Western blot analysis revealed that the rat optic nerve showed immunoreactivity to antibodies against GLT-1, GLAST, and EAAC1. In conclusion, we suggest that glial and neuronal transporters are present in the rat optic nerve, where dynamic axon-glial interaction has been known to occur. In particular, the unusually high level of expression of GLAST in the optic nerve suggests a possible role for this glial transporter in protecting optic nerves from neurotoxicity during postnatal development. GLIA 20:184–192, 1997. © 1997 Wiley-Liss, Inc.     

Effects of cue exposure on brain glucose utilization 8 days after repeated cocaine administration

Relapse to cocaine use may involve exposure to cocaine-associated environmental cues. The present experiment tested the hypothesis that basal local cerebral metabolic rate for glucose (LCMR(glu)), as measured by the 2-deoxy-D-[l-(14)C]glucose (2-DG) autoradiography, would change in the presence of cocaine conditioned cues at 8 days after the last of seven daily cocaine injections (30 mg/kg). This dose regimen results in sensitization to the locomotor effects of cocaine. Cocaine was administered to two groups of rats while saline was administered to a third. In the conditioned group, the rats were placed into the 2-DG experimental chamber immediately after cocaine injection. Rats in the non-conditioned group were placed into their home cage after cocaine administration. A control group received only saline. The 2-DG experiment was conducted in non-drugged animals 8 days after treatment completion. The interaction between treatment status and brain region was significant. Mean basal LCMR(glu) was significantly lower in 12 brain regions in the conditioned group as compared to the control group, but was significantly lower in only four areas in the non-conditioned group. Regions in which there were significant changes in the conditioned group included the basolateral amygdala, subiculum, medial thalamus, lateral habenula and the substantia nigra pars compacta. LCMR(glu) was significantly reduced in the ventrolateral orbital cortex and rostral nucleus accumbens in both experimental groups. These findings indicate that repeated cocaine administration can cause protracted decreases in basal LCMR(glu), decreases that are more widespread in the brain during exposure to cocaine-associated cues.     

Gene expression of glucose transporters and glycolytic enzymes in the CNS of rats behaviorally dependent on ethanol

The steady-state levels of messenger RNA (mRNA) of the glucose transporters 1 and 3 and the glycolytic enzymes hexokinase, phosphofructokinase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate dehydrogenase were measured in up to seven brain regions of the rat in a recently developed animal model of 'behavioral dependence' on ethanol. Irreversible behavioral dependence, including loss of control, was induced by offering the rats the choice between ethanol and water over a 9-month period (Group A). This group was compared with a group given the choice between ethanol and water for only 2 months (not yet behaviorally dependent, Group B), a group forced to consume ethanol as sole fluid over a 9-month period (not behaviorally dependent, Group C) and ethanol-naive control rats. All groups were sacrificed 1 month after ethanol withdrawal. The mRNA concentrations of both neuronal glucose transporter 3 and the key glycolytic enzymes phosphofructokinase and pyruvate dehydrogenase were significantly reduced in the hippocampi of the rats behaviorally dependent on ethanol (Group A). No significant changes were seen in any of the remaining brain regions (e.g., cortical areas, limbic forebrain, amygdala, midbrain) in Group A, or in any brain area at all in Groups B and C. The results show that chronic consumption of ethanol in a free-choice situation may impair neuronal glucose uptake and glycolytic flux. This effect is manifested exclusively in the hippocampus and is specifically related to the development of behavioral dependence, since it was not found after forced administration of large amounts of ethanol (Group C).     

Loss of glial glutamate transporters and induction of neuronal expression of GLT-1B in the hypoxic neonatal pig brain

The homeostasis of glutamate is critical to normal brain function; deficiencies in the regulation of extracellular glutamate are thought to be a major determinant of damage in hypoxic brains. Extracellular levels of glutamate are regulated mainly by plasmalemmal glutamate transporters. We have evaluated the distribution of the glutamate transporter GLAST and two splice variants of GLT-1 in the hypoxic neonatal pig brain using this as model of neonatal humans. In response to severe hypoxic insults, we observe a rapid loss of two glial glutamate transporters from specific brain regions, such as the CA1 region of the hippocampus, but not the dentate gyrus. The spatial distribution of loss accords with patterns of damage in these brains. Conversely, we demonstrate that hypoxia evokes the expression of a splice variant of GLT-1 in neurons. We suggest that this expression may be induced in response to elevated extracellular glutamate around these neurons, and that this splice variant may represent a useful marker for direct quantification of the extent of likely neuronal damage in hypoxic brains.     

Consequences of amygdala kindling and repeated withdrawal from ethanol on amphetamine-induced behaviours

It has been shown previously that chronic ethanol treatment in mice leads to accelerated behavioural sensitization to psychomotor stimulants [Manley & Little (1997) J. Pharmacol. Exp. Ther., 281, 1330-1339], whilst repeated experience of ethanol withdrawal sensitizes pathways underlying seizure activity (Becker & Hale (1993) Alcohol Clin. Exp. Res., 17, 94-98]. The aim of the current experiment was to investigate the consequences of repeated withdrawal from ethanol on amphetamine-induced behaviours in the rat and compare this with animals with electrical kindling of the amygdala, a procedure that has been shown to enhance alcohol withdrawal seizures [Pinel et al. (1975) Can. J. Neurol. Sci., 2, 467-475]. For the kindling experiments, electrodes were surgically implanted in the left basolateral amygdala and were stimulated daily at the afterdischarge threshold until a criterion of three consecutive stage 5 seizures was reached. Fully kindled rats showed a marginally significant reduction in sensitivity to the locomotor stimulant effects of acute amphetamine compared with sham and partially kindled rats which had experienced subthreshold stimulation of the amygdala. Sham and partially kindled rats sensitized readily to the locomotor activating effects of amphetamine (0.125 mg/kg) following repeated treatments, but the fully kindled rats did not. Fully kindled rats also failed to show place preference conditioning to amphetamine (0.5 mg/kg). Rats, withdrawn three times from chronic ethanol (liquid-diet), kindled more quickly to PTZ (30 mg/kg, i.p.) than rats with the same overall exposure to ethanol (24 days) followed by a single withdrawal or control animals. However, there was no difference in the locomotor stimulating effects of acute amphetamine (0.25-1 mg/kg, i.p.), the rate of sensitization to amphetamine (0.125 mg/kg, i.p.) or amphetamine induced conditioned place preference (1 mg/kg, i.p.). These observations suggest that, in rats, repeated withdrawal from a relatively mild chronic ethanol treatment modulates neuronal systems that may also be involved in PTZ-induced kindling but not those involved in either the acute stimulant effects of amphetamine or behavioural sensitization or appetitive conditioning following repeated amphetamine administration. Behavioural changes following amygdala kindling differed from those following repeated ethanol withdrawal, suggesting that withdrawal kindling from a mild ethanol treatment differs in its effects from amygdala kindling.     

Role of glial glutamate transporters in the facilitatory action of FK960 on hippocampal neurotransmission.

We found previously that N-(4-acetyl-1-piperazinyl)-p-fluorobenzamide monohydrate (FK960) facilitated hippocampal neurotransmission in the dentate gyrus of rat hippocampal slices. The present study was conducted to understand the mechanism underlying the facilitatory action of FK960. The facilitation was inhibited by H-89, an inhibitor of cAMP-dependent protein kinase (PKA), but it was not affected by cycloheximide, a protein synthesis blocker. In cultured rat hippocampal neurons, the drug had no effect on either spontaneous miniature excitatory postsynaptic currents or whole-cell membrane currents evoked by glutamate, kainate, or NMDA, suggesting that the facilitatory action of FK960 is not caused by increasing presynaptic transmitter release or excitatory postsynaptic conductances. FK960 inhibited responses of the glial glutamate transporter, GLT-1, expressed in Xenopus oocytes, and a similar effect was found with cultured rat astrocytes. The FK960 action was inhibited in the presence of H-89. The results of the present study thus suggest that FK960 facilitates hippocampal neurotransmission by inhibiting GLT-1 glial glutamate reuptake via a PKA pathway, thereby increasing synaptic glutamate concentrations.