Lower Glutamate Levels Research Articles

From Allostatic Load to Allostatic State-An Endogenous Sympathetic Strategy to Deal With Chronic Anxiety and Stress?

The concepts of allostatic load and overload, i. e., a dramatic increase in the allostatic load that predisposes to disease, have been extensively described in the literature. Here, we show that rats engaging in active offensive response (AOR) behavioral strategies to chronic predator scent stress (PSS) display less anxiety behavior and lower plasma cortisol levels vs. rats engaging in passive defensive response (PDR) behavioral strategies to chronic PSS. In the same chronic PSS paradigm, AOR rats also have higher lactate and lower glutamate levels in amygdala but not in control-region hippocampus vs. PDR rats. The implications of these findings for regulation of allostatic and stress responses, and post-traumatic stress disorder (PTSD) are discussed.

Assessing Brain Metabolism With 7-T Proton Magnetic Resonance Spectroscopy in Patients With First-Episode Psychosis

The use of high-field magnetic resonance spectroscopy (MRS) in multiple brain regions of a large population of human participants facilitates in vivo study of localized or diffusely altered brain metabolites in patients with first-episode psychosis (FEP) compared to healthy participants. To compare metabolite levels in 5 brain regions between patients with FEP (evaluated within 2 years of onset) and healthy controls, and to explore possible associations between targeted metabolite levels and neuropsychological test performance. Cross-sectional design used 7-T MRS at a research MR imaging facility in participants recruited from clinics at the Johns Hopkins Schizophrenia Center and the local population. Eighty-one patients who had received a DSM-IV diagnosis of FEP within the last 2 years and 91 healthy age-matched (but not sex-matched) volunteers participated. Brain metabolite levels including glutamate, glutamine, γ-aminobutyric acid (GABA), N-acetylaspartate, N-acetylaspartyl glutamate, and glutathione, as well as performance on neuropsychological tests. The mean (SD) age of 81 patients with FEP was 22.3 (4.4) years and 57 were male, while the mean (SD) age of 91 healthy participants was 23.3 (3.9) years and 42 were male. Compared with healthy participants, patients with FEP had lower levels of glutamate (F1,162 = 8.63, P = .02), N-acetylaspartate (F1,161 = 5.93, P = .03), GABA (F1,163 = 6.38, P = .03), and glutathione (F1,162 = 4.79, P = .04) in the anterior cingulate (all P values are corrected for multiple comparisons); lower levels of N-acetylaspartate in the orbitofrontal region (F1,136 = 7.23, P = .05) and thalamus (F1,133 = 6.78, P = .03); and lower levels of glutathione in the thalamus (F1,135 = 7.57, P = .03). Among patients with FEP, N-acetylaspartate levels in the centrum semiovale white matter were significantly correlated with performance on neuropsychological tests, including processing speed (r = 0.48; P < .001), visual (r = 0.33; P = .04) and working (r = 0.38; P = .01) memory, and overall cognitive performance (r = 0.38; P = .01). Seven-tesla MRS offers insights into biochemical changes associated with FEP and may be a useful tool for probing brain metabolism that ranges from neurotransmission to stress-associated pathways in participants with psychosis.

Glutamine has antidepressive effects through increments of glutamate and glutamine levels and glutamatergic activity in the medial prefrontal cortex

Emerging evidence has shown the low levels of glutamate (Glu) and glutamine (Gln) and the hypoactivity in the cortex of patients with depression. The hypoactivity is closely related with low frequency of glutamatergic signaling that is affected by the levels of Glu and Gln. Thus, we hypothesized that there might be a causality among low levels of Glu and Gln, hypoactive glutamatergic neurotransmissions, and depressive behaviors. Here, we found low Glu and Gln levels and low frequency of spontaneous excitatory postsynaptic current (sEPSC) of glutamatergic neurons in the medial prefrontal cortex (mPFC) of chronic immobilization stress (CIS)-induced depressed mice. The depressed mice also showed hypoactive Gln synthetase (GS). Inhibition of GS by methionine sulfoximine (MSO) decreased Glu and Gln levels and increased depressive behaviors with low frequency of sEPSC in the mPFC, indicating that Glu and Gln decrements cause hypoactive glutamatergic neurotransmissions and depressive behaviors. Both Glu and Gln could increase sEPSC of glutamatergic neurons in the mPFC on slice patch, but only Gln overcame MSO to increase sEPSC, suggesting that exogenous Gln would recover CIS-induced low frequency of sEPSC caused by hypoactive GS and act as an antidepressant. Expectedly, Gln supplementation showed antidepressant effects against CIS; it increased glutamatergic neurotransmissions with Glu and Gln increment in the mPFC and attenuated depressive behaviors. Moreover, selective glutamatergic activation in the mPFC by optogenetics decreased depressive behavior. In conclusion, depressive behaviors evoked by chronic stress were due to hypoactive glutamatergic neurons in the mPFC caused by low levels of Glu and Gln, and exogenous Gln can be used as an alternative antidepressant to increase glutamatergic neurotransmission.

25.2 UNDERSTANDING WHITE MATTER PATHOLOGY IN SCHIZOPHRENIA USING DIFFUSION TENSOR IMAGING AND MAGNETIC RESONANCE SPECTROSCOPY

BackgroundDiffusion tensor imaging (DTI) studies in schizophrenia consistently show global reductions in fractional anisotropy (FA), a putative marker of white matter integrity. Because magnetic resonance spectroscopy (MRS) studies permit for the non-invasive measurements of neurometabolites, such as N-acetylaspartate (NAA), a marker of neuronal integrity, and glutamate, which can be neurotoxic when over-released, this technique provides further understanding of the relationship between white matter microstructure and neuronal function.MethodsTwenty-nine schizophrenia patients and twenty controls participated in this 3T imaging study where we used DTI and tract-based spatial statistics (TBSS) to assess white matter integrity of the cingulum bundle and MRS to quantify NAA and glutamate in the anterior cingulate cortex (ACC) and hippocampus, i.e. in cortico-limbic regions connected by the cingulum bundle.ResultsWe found FA reductions with overlapping radial diffusivity (RD) elevations in patients in multiple tracts, suggesting white matter abnormalities in schizophrenia are driven by loss of myelin integrity. In controls, but not in patients, high hippocampal NAA levels were significantly associated with low RD in the hippocampal part of the cingulum, and low ACC glutamate levels were significantly associated with high FA in the hippocampus part of the cingulum.DiscussionIn conclusion, we demonstrate the potential utility of a multi-modal neuroimaging approach to help further our understanding of the relationship between white matter microstructure and neurochemistry in distinct cortical regions connected by white matter tracts.

Glutaminolysis drives membrane trafficking to promote invasiveness of breast cancer cells

The role of glutaminolysis in providing metabolites to support tumour growth is well-established, but the involvement of glutamine metabolism in invasive processes is yet to be elucidated. Here we show that normal mammary epithelial cells consume glutamine, but do not secrete glutamate. Indeed, low levels of extracellular glutamate are necessary to maintain epithelial homoeostasis, and provision of glutamate drives disruption of epithelial morphology and promotes key characteristics of the invasive phenotype such as lumen-filling and basement membrane disruption. By contrast, primary cultures of invasive breast cancer cells convert glutamine to glutamate which is released from the cell through the system Xc- antiporter to activate a metabotropic glutamate receptor. This contributes to the intrinsic aggressiveness of these cells by upregulating Rab27-dependent recycling of the transmembrane matrix metalloprotease, MT1-MMP to promote invasive behaviour leading to basement membrane disruption. These data indicate that acquisition of the ability to release glutamate is a key watershed in disease aggressiveness.

Intelligence and Brain Efficiency: Investigating the Association between Working Memory Performance, Glutamate, and GABA.

Intelligence is a measure of general cognitive functioning capturing a wide variety of different cognitive functions. It has been hypothesized that the brain works to minimize the resources allocated toward higher cognitive functioning. Thus, for the intelligent brain, it may be that not simply more is better, but rather, more efficient is better. Energy metabolism supports both inhibitory and excitatory neurotransmission processes. Indeed, in glutamatergic and GABAergic neurons, the primary energetic costs are associated with neurotransmission. We tested the hypothesis that minimizing resources through the excitation–inhibition balance encompassing gamma-aminobutyric acid (GABA) and glutamate may be beneficial to general cognitive functioning using 7 T 1H-MRS in 23 healthy individuals (male/female = 16/7, 27.7 ± 5.3 years). We find that a higher working memory index is significantly correlated with a lower GABA to glutamate ratio in the frontal cortex and with a lower glutamate level in the occipital cortex. Thus, it seems that working memory performance is associated with the excitation–inhibition balance in the brain.

Unique prefrontal GABA and glutamate disturbances in co-occurring bipolar disorder and alcohol dependence

Bipolar disorder (BD) and alcohol dependence (AD) frequently co-occur, and co-occurring BD and AD are associated with devastating public health costs. Minimal neurobiological research exists to guide the development of effective treatments for this treatment-resistant population. We believe the present study represents the first investigation of prefrontal gamma-aminobutyric acid (GABA) and glutamate levels in co-occurring BD and current AD. The participants were 78 individuals who met DSM-IV criteria for BD I/II and current AD (n=20), BD I/II alone (n=19), current AD alone (n=20) or no diagnosis (n=19). The participants completed a baseline diagnostic visit, then returned approximately 4 days later for a two-dimensional J-resolved proton magnetic resonance spectroscopy (1H-MRS) acquisition in dorsal anterior cingulate cortex (dACC). All participants were required to demonstrate ⩾1 week of abstinence from alcohol/drugs via serial biomarker testing before 1H-MRS. A 2 × 2 factorial analysis of variance of cerebrospinal fluid (CSF)-corrected GABA/water concentrations demonstrated a significant BD × AD interaction (F=2.91, P<0.05), signifying uniquely low levels of GABA in BD+AD; this effect doubled when the sample was restricted to individuals who consumed alcohol within 2 weeks of 1H-MRS. There were no overall effects of BD/AD on CSF-corrected glutamate/water levels. However, the BD × AD interaction, signifying uniquely low levels of glutamate in BD+AD, approached statistical significance (F=3.83, P=0.06) in individuals who consumed alcohol within 2 weeks of 1H-MRS. The dACC GABA levels were significantly, negatively associated with Barratt Impulsiveness Scale (r=−0.28, P=0.02) and Obsessive Compulsive Drinking Scale (r=−0.35, P<0.01) scores. If replicated, these results may suggest that future treatment studies should preferentially evaluate therapeutics in BD+AD known to increase prefrontal GABA and glutamate levels.

Effects of Antipsychotic Administration on Brain Glutamate in Schizophrenia: A Systematic Review of Longitudinal 1H-MRS Studies.

Schizophrenia is associated with brain glutamate dysfunction, but it is currently unclear whether antipsychotic administration can reduce the extent of glutamatergic abnormality. We conducted a systematic review of proton magnetic resonance spectroscopy (1H-MRS) studies examining the effects of antipsychotic treatment on brain glutamate levels in schizophrenia. The Medline database was searched to identify relevant articles published until December 2016. Inclusion required that studies examined longitudinal changes in brain glutamate metabolites in patients with schizophrenia before and after initiation of first antipsychotic treatment or a switch in antipsychotic treatment. The searches identified eight eligible articles, with baseline and follow-up measures in a total of 168 patients. The majority of articles reported a numerical reduction in brain glutamate metabolites with antipsychotic treatment, and the estimated overall mean reduction of 6.5% in Glx (the combined signal from glutamate and glutamine) across brain regions. Significant reductions in glutamate metabolites in at least one brain region were reported in four of the eight studies, and none of the studies reported a significant glutamatergic increase after antipsychotic administration. Relationships between the degree of change in glutamate and the degree of improvement in symptoms have been inconsistent but may provide limited evidence that antipsychotic response may be associated with lower glutamate levels before treatment and a greater extent of glutamatergic reduction during treatment. Further longitudinal, prospective studies of glutamate and antipsychotic response are required to confirm these findings.

4.1 Glutamatergic Alterations Across the Schizophrenia Illness Course

Abstract Background: Glutamate may decline with age (or illness course) in schizophrenia at a greater rate than the psychiatrically healthy population as suggested by a meta-analysis. However, no studies have specifically addressed this question. Therefore, we investigated glutamate levels with magnetic resonance spectroscopy (MRS), in younger and older participants with schizophrenia in 2 separate studies with differing samples and methods. Regions strongly implicated in the pathophysiology of schizophrenia-related disorders, the anterior cingulate and the medial temporal lobe, were investigated. Methods: MR scanning was conducted on a 3T Siemens Tim Trio equipped with a 32-channel head coil. One hundred and sixty-seven participants completed Study 1, and 86 participants completed Study 2. For Study 1, spectra were acquired from the anterior cingulate using phase rotation STEAM (PR-STEAM) (13,14): TR/TM/TE = 2000/10/6.5 ms, NEX = 128, and phases: φ1 = 135°, φ2 = 22.5°, φ13 = 112.5°, φADC = 0° and a water reference (NEX = 16). Glutamate and glutamine were quantified. For Study 2, spectra were acquired from the medial temporal lobe (i.e., hippocampus) and parietal lobe using a PRESS sequence (TR/TE = 2000, 30 ms, NEX = 256 for hippocampus and 128 for parietal and a water reference NEX = 16). Patients were evaluated for psychopathology, and all participants completed cognitive testing with a focus on attention and working memory for Study 1 and relational memory for Study 2. Results: For Study 1, results revealed significant age (P < .001) and diagnosis (P = .001) effects for glutamate such that glutamate was lower in participants with schizophrenia compared to controls and in older versus younger participants. There was also a significant age effect (P = .007) for glutamine such that glutamine levels increased with age in both the groups. Higher anterior cingulate glutamate was related to better attention (P < .001) and working memory (P = .04). For Study 2, hippocampal glutamate was lower in older but not younger participants with schizophrenia (P = .03, Diagnosis × Age interaction). Parietal glutamate was lower in participants with schizophrenia irrespective of age (P = .013). Better relational memory performance was related to higher levels of parietal glutamate (P < .01). Conclusion: Participants with schizophrenia were distinguished by lower anterior cingulate and parietal glutamate levels, but there was no evidence of accelerated glutamate aging in these brain regions. Lower hippocampal glutamate levels observed in older participants with schizophrenia suggests that the hippocampus may be particularly vulnerable to aging in schizophrenia. Glutamatergic interventions targeted at increasing levels in the anterior cingulate and parietal cortex and halting the decline in the hippocampus may improve cognitive function and quality of life in patients with schizophrenia.

Further studies of the effects of aging on arginine metabolites in the rat vestibular nucleus and cerebellum

Some studies have demonstrated that aging is associated with impaired vestibular reflexes, especially otolithic reflexes, resulting in postural instability. However, the neurochemical basis of these age-related changes is still poorly understood. The l-arginine metabolic system has been implicated in changes in the brain associated with aging. In the current study, we examined the levels of l-arginine and its metabolizing enzymes and downstream metabolites in the vestibular nucleus complex (VNC) and cerebellum (CE) of rats with and without behavioral testing which were young (4months old), middle-aged (12months old) or aged (24months old). We found that aging was associated with lower nitric oxide synthase activity in the CE of animals with testing and increased arginase in the VNC and CE of animals with testing. l-citrulline and l-ornithine were lower in the VNC of aged animals irrespective of testing, while l-arginine and l-citrulline were lower in the CE with and without testing, respectively. In the VNC and CE, aging was associated with lower levels of glutamate in the VNC, irrespective of testing. In the VNC it was associated with higher levels of agmatine and putrescine, irrespective of testing. In the CE, aging was associated with higher levels of putrescine in animals without testing and with higher levels of spermine in animals with testing, and spermidine, irrespective of testing. Multivariate analyses indicated significant predictive relationships between the different variables, and there were correlations between some of the neurochemical variables and behavioral measurements. Cluster analyses revealed that aging altered the relationships between l-arginine and its metabolites. The results of this study demonstrate that there are major changes occurring in l-arginine metabolism in the VNC and CE as a result of age, as well as behavioral activity.

Constitutive regulation of the glutamate/aspartate transporter EAAT1 by Calcium-Calmodulin-Dependent Protein Kinase II.

Glutamate clearance by astrocytes is an essential part of normal excitatory neurotransmission. Failure to adapt or maintain low levels of glutamate in the central nervous system is associated with multiple acute and chronic neurodegenerative diseases. The primary excitatory amino acid transporters in human astrocytes are EAAT1 and EAAT2 (GLAST and GLT-1, respectively, in rodents). While the inhibition of calcium/calmodulin-dependent kinase (CaMKII), a ubiquitously expressed serine/threonine protein kinase, results in diminished glutamate uptake in cultured primary rodent astrocytes (Ashpole etal. 2013), the molecular mechanism underlying this regulation is unknown. Here, we use a heterologous expression model to explore CaMKII regulation of EAAT1 and EAAT2. In transiently transfected HEK293T cells, pharmacological inhibition of CaMKII (using KN-93 or tat-CN21) reduces [3 H]-glutamate uptake in EAAT1 without altering EAAT2-mediated glutamate uptake. While over-expressing the Thr287Asp mutant to enhance autonomous CaMKII activity had no effect on either EAAT1 or EAAT2-mediated glutamate uptake, over-expressing a dominant-negative version of CaMKII (Asp136Asn) diminished EAAT1 glutamate uptake. SPOTS peptide arrays and recombinant glutathione S-transferase-fusion proteins of the intracellular N- and C-termini of EAAT1 identified two potential phosphorylation sites at residues Thr26 and Thr37 in the N-terminus. Introducing an Ala (a non-phospho mimetic) at Thr37 diminished EAAT1-mediated glutamate uptake, suggesting that the phosphorylation state of this residue is important for constitutive EAAT1 function. Our study is the first to identify a glutamate transporter as a direct CaMKII substrate and suggests that CaMKII signaling is a critical driver of constitutive glutamate uptake by EAAT1.

Effect of Bilingualism in the Anterior Cingulate Cortex: A Proton Magnetic Resonance Spectroscopy Study in Two Centres

Effect of Bilingualism in the Anterior Cingulate Cortex: A Proton Magnetic Resonance Spectroscopy Study in Two Centres

The dose makes the poison: from glutamate-mediated neurogenesis to neuronal atrophy and depression.

Experimental evidence has demonstrated that glutamate is an essential factor for neurogenesis, whereas another line of research postulates that excessive glutamatergic neurotransmission is associated with the pathogenesis of depression. The present review shows that such paradox can be explained within the framework of hormesis, defined as biphasic dose responses. Low glutamate levels activate adaptive stress responses that include proteins that protect neurons against more severe stress. Conversely, abnormally high levels of glutamate, resulting from increased release and/or decreased removal, cause neuronal atrophy and depression. The dysregulation of the glutamatergic transmission in depression could be underlined by several factors including a decreased inhibition (γ-aminobutyric acid or serotonin) or an increased excitation (primarily within the glutamatergic system). Experimental evidence shows that the activation of N-methyl-D-aspartate receptor (NMDA) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors (AMPAR) can exert two opposite effects on neurogenesis and neuron survival depending on the synaptic or extrasynaptic concentration. Chronic stress, which usually underlies experimental and clinical depression, enhances glutamate release. This overactivates NMDA receptors (NMDAR) and consequently impairs AMPAR activity. Various studies show that treatment with antidepressants decreases plasma glutamate levels in depressed individuals and regulates glutamate receptors by reducing NMDAR function by decreasing the expression of its subunits and by potentiating AMPAR-mediated transmission. Additionally, it has been shown that chronic treatment with antidepressants having divergent mechanisms of action (including tricyclics, selective serotonin reuptake inhibitors, and ketamine) markedly reduced depolarization-evoked glutamate release in the hippocampus. These data, taken together, suggest that the glutamatergic system could be a final common pathway for antidepressant treatments.

Frontal Glutamate and γ-Aminobutyric Acid Levels and Their Associations With Mismatch Negativity and Digit Sequencing Task Performance in Schizophrenia.

Auditory mismatch negativity (MMN) is a biomarker for schizophrenia thought to reflect glutamatergic N-methyl-d-aspartate receptor function and excitatory-inhibitory neurotransmission balance. However, the association of glutamate level with MMN has not been directly examined in patients with schizophrenia, to our knowledge. To investigate the contributions of glutamate and γ-aminobutyric acid (GABA) to MMN and digit sequencing task (DST) performance, an assessment of verbal working memory, in schizophrenia. Fifty-three control participants from the community and 45 persons with schizophrenia from outpatient clinics completed an electroencephalographic session for MMN, magnetic resonance spectroscopy for glutamate and GABA, and a DST. The study dates were July 2011 to May 2014, and the dates of our analysis were May 2014 to August 2015. Glutamate, GABA, the ratio of glutamine to glutamate, MMN amplitude, and DST. Structural equation modeling was used to test the effects of neurochemistry and MMN amplitude on DST performance. The 45 persons with schizophrenia were a mean (SD) of 37.7 (12.8) years and the control participants were 37.1 (13.1) years. The schizophrenia group had a mean (SD) of 14.7 (12.1) years of illness. Mismatch negativity amplitude (F = 4.39, P = .04) and glutamate (F = 9.69, P = .002) were reduced in the schizophrenia group. Smaller MMN amplitude was significantly associated with lower GABA level (P = .008), lower glutamate level (P = .05), and higher ratio of glutamine to glutamate (P = .003). Reduced MMN amplitude was linked to poor verbal working memory in schizophrenia (P = .002). Modeling revealed that a proxy of glutamatergic function, indexed by the ratio of glutamine to glutamate, influenced a path from the ratio of glutamine to glutamate to MMN to verbal working memory (P = .38 [root-mean-square error of approximation, P < .001] by χ2 test), supporting the contention that MMN serves as an intermediate biomarker linking glutamatergic function to DST performance in schizophrenia. The role of glutamate and GABA in MMN and verbal working memory deficits in schizophrenia has been frequently debated. These data provide in vivo evidence that support glutamatergic and GABAergic regulation of MMN and verbal working memory function in schizophrenia.

Cardiac Resynchronization Therapy Induces Adaptive Metabolic Transitions in the Metabolomic Profile of Heart Failure

BackgroundHeart failure (HF) is associated with ventricular dyssynchrony and energetic inefficiency, which can be alleviated by cardiac resynchronization therapy (CRT). The aim of this study was to determine the metabolomic signature in HF and its prognostic value regarding the response to CRT. Methods and ResultsThis prospective study consisted of 24 patients undergoing CRT for advanced HF and 10 control patients who underwent catheter ablation for supraventricular arrhythmia but not CRT. Blood samples were collected before and 3 months after CRT. Metabolomic profiling of plasma samples was performed with the use of gas chromatography–mass spectrometry and nuclear magnetic resonance. The plasma metabolomic profile was altered in the HF patients, with a distinct panel of metabolites, including Krebs cycle and lipid, amino acid, and nucleotide metabolism. CRT improved the metabolomic profile. The succinate-glutamate ratio, an index of Krebs cycle activity, improved from 0.58 ± 0.13 to 2.84 ± 0.60 (P < .05). The glucose-palmitate ratio, an indicator of the balance between glycolytic and fatty acid metabolism, increased from 0.96 ± 0.05 to 1.54 ± 0.09 (P < .01). Compared with nonresponders to CRT, responders had a distinct baseline plasma metabolomic profile, including higher isoleucine, phenylalanine, leucine, glucose, and valine levels and lower glutamate levels at baseline (P < .05). ConclusionsCRT improves the plasma metabolomic profile of HF patients, indicating harmonization of myocardial energy substrate metabolism. CRT responders may have a favorable metabolomic profile as a potential biomarker for predicting CRT outcome.

Up-Regulation of TREK-2 Potassium Channels in Cultured Astrocytes Requires De Novo Protein Synthesis: Relevance to Localization of TREK-2 Channels in Astrocytes after Transient Cerebral Ischemia

Excitotoxicity due to glutamate receptor over-activation is one of the key mediators of neuronal death after an ischemic insult. Therefore, a major function of astrocytes is to maintain low extracellular levels of glutamate. The ability of astrocytic glutamate transporters to regulate the extracellular glutamate concentration depends upon the hyperpolarized membrane potential of astrocytes conferred by the presence of K+ channels in their membranes. We have previously shown that TREK-2 potassium channels in cultured astrocytes are up-regulated by ischemia and may support glutamate clearance by astrocytes during ischemia. Thus, herein we determine the mechanism leading to this up-regulation and assess the localization of TREK-2 channels in astrocytes after transient middle cerebral artery occlusion. By using a cell surface biotinylation assay we confirmed that functional TREK-2 protein is up-regulated in the astrocytic membrane after ischemic conditions. Using real time RT-PCR, we determined that the levels of TREK-2 mRNA were not increased in response to ischemic conditions. By using Western blot and a variety of protein synthesis inhibitors, we demonstrated that the increase of TREK-2 protein expression requires De novo protein synthesis, while protein degradation pathways do not contribute to TREK-2 up-regulation after ischemic conditions. Immunohistochemical studies revealed TREK-2 localization in astrocytes together with increased expression of the selective glial marker, glial fibrillary acidic protein, in brain 24 hours after transient middle cerebral occlusion. Our data indicate that functional TREK-2 channels are up-regulated in the astrocytic membrane during ischemia through a mechanism requiring De novo protein synthesis. This study provides important information about the mechanisms underlying TREK-2 regulation, which has profound implications in neurological diseases such as ischemia where astrocytes play an important role.

Astrocytes in the medial preoptic area modulate ejaculation latency in an experience-dependent fashion.

While sexually experienced males copulate at a higher frequency than sexually inexperienced males, there is still a great deal of variability in their behavior. Within the medial preoptic area (mPOA) of the hypothalamus, glutamate modulates some of this variability. Glutamate levels, for example, increase during sexual activity, peaking with ejaculation and falling precipitously during the post-ejaculation interval. Whereas lower glutamate levels after ejaculation translates to longer post-ejaculatory intervals, administration of glutamate uptake inhibitors into the mPOA increases the number of ejaculations a male rat achieves over a mating bout, and reduces the latency to ejaculate once mating begins. Because astrocytes modulate the availability of neuronal glutamate, we hypothesized that differences in the number of GFAP-positive astrocytes in the mPOA may account for variability in sexual behavior. To this end, we examined whether the number of astrocytes in the mPOA related to ejaculation latency as well as to the duration of the post-ejaculatory interval (PEI) in sexually experienced and sexually inexperienced males. Results indicate that the number of astrocytes negatively correlated with latency to reach ejaculations in sexually inexperienced but not sexually experienced rats while the number of astrocytes and PEI were not related. Astrocyte numbers did not vary between inexperienced and experienced subjects indicating that astrocyte processes may differentially project to sex-relevant glutamatergic synapses or that glutamatergic innervation of the mPOA changes as a function of sexual experience.

Loss of SIRT4 decreases GLT-1-dependent glutamate uptake and increases sensitivity to kainic acid.

Glutamate transport is a critical process in the brain that maintains low extracellular levels of glutamate to allow for efficient neurotransmission and prevent excitotoxicity. Loss of glutamate transport function is implicated in epilepsy, traumatic brain injury, and amyotrophic lateral sclerosis. It remains unclear whether or not glutamate transport can be modulated in these disease conditions to improve outcome. Here, we show that sirtuin (SIRT)4, a mitochondrial sirtuin, is up-regulated in response to treatment with the potent excitotoxin kainic acid. Loss of SIRT4 leads to a more severe reaction to kainic acid and decreased glutamate transporter expression and function in the brain. Together, these results indicate a critical and novel stress response role for SIRT4 in promoting proper glutamate transport capacity and protecting against excitotoxicity.

METABOLIC IMAGING BIOMARKERS FOR MUTANT IDH1 GLIOMAS

BACKGROUND: The IDH1 mutation is now recognized as an important driver in the etiology of low-grade brain tumors. In contrast to wild-type IDH1, which catalyzes the conversion of isocitrate to α-ketoglutarate (αKG), mutant IDH1 converts αKG into the oncometabolite 2-hydroxygluatrate (2HG). In an effort to noninvasively image the activity of mutant IDH1, we have previously shown that we can use hyperpolarized αKG together with 13C magnetic resonance spectroscopy (MRS) to noninvasively detect the production of hyperpolarized 2HG in vivo. In more recent studies we have focused on identifying additional metabolic alterations associated with the presence of the IDH1 mutation in order to develop complementary metabolic imaging approaches that might serve to inform on the presence of mutant IDH1 and its modulation with targeted therapies. METHODS: Two mutant and wild-type IDH1 cell models were generated by using either U87 glioblastoma cells or E6/E7/hTERT immortalized normal human astrocytes (NHAs) and lentivirally transducing with either wild-type IDH1 (wt cells) or wild-type and mutant IDH1 (mutant cells). Using 1H MRS we investigated cell extracts to determine steady state metabolite levels. We also determined metabolic fluxes by using 13C MRS and probing the fate of 13C-labeled precursors (at thermal equilibrium or hyperpolarized) in live cells grown in a bioreactor or in orthotopic tumors in rats. RESULTS: We found that in both U87 and NHA-based cell models not only was the IDH1 mutation associated with elevated levels of 2HG, but other metabolite levels were also altered. In particular, steady state glutamate levels were lower. This was associated with an increased flux of glutamine to 2-HG and a concomitantly reduced flux of glutamine to glutamate. The flux of glucose into the TCA cycle and towards glutamate was also reduced. At the same time the activity and expression of BCAT1 and aspartate aminotransferase dropped. Pyruvate dehydrogenase activity was also reduced, associated with an increase in its phosphorylation and the expression of pyruvate dehydrogenase kinase 3. Collectively our findings likely explain the lower glutamate levels observed in mutant IDH1 cells. Importantly, we were also able to detect a drop in the potentially translatable in vivo imaging of hyperpolarized glutamate produced from hyperpolarized αKG in mutant IDH1 orthotopic tumors. CONCLUSIONS: In addition to the production of 2HG, the presence of mutant IDH1 leads to MRS-detectable metabolic changes that could serve as complementary imaging biomarkers of mutant IDH1 activity or its inhibition with treatment. SECONDARY CATEGORY: n/a.

Oxidative stress increases in retinas of dogs in acute glaucoma but not in chronic glaucoma.

To test the hypothesis that oxidative stress occurs early in the pathogenesis of glaucoma in dogs. Sections from eight control retinas and 25 retinas from dogs with primary glaucoma. For retinas embedded in paraffin, sections were immunohistochemically stained for malondialdehyde (MDA) and 3-nitrotyrosine (NT). For retinas embedded in plastic, serial 0.5-μm sections were immunogold-stained for total glutathione, taurine, and glutamate. Increased immunostaining for MDA and NT, markers of oxidative stress, occurred in retinal ganglion cells (RGCs) and other neurons in acute glaucoma, but not in chronic glaucoma. In minimally damaged regions, immunostaining for the antioxidant glutathione was decreased in RGCs, neurons of the inner nuclear layer (INL), and Müller cell processes. The loss of glutathione immunostaining in RGCs occurred without a decrease in glutamate immunostaining. Neurons with nuclear damage in the INL had low levels of glutathione, taurine, and glutamate. In severely damaged regions, immunostaining for glutathione was increased in the remaining retinal tissue. Immunohistochemical staining revealed an increase in markers of oxidative stress and loss of glutathione in neurons with minimal damage during acute glaucoma. Oxidative changes were no longer present in chronic glaucomatous retinas, suggesting transient oxidative stress occurs early in glaucoma. The loss of glutathione in minimally damaged regions occurred without a significant redistribution of glutamate, suggesting oxidative stress may occur before glutamate redistribution. Alteration in markers of oxidative stress occurs early in canine glaucoma, suggesting oxidative stress may contribute to subsequent glutamate redistribution and other damaging processes.