Affinage

SLC1A2

Excitatory amino acid transporter 2 · UniProt P43004

Length
574 aa
Mass
62.1 kDa
Annotated
2026-04-28
100 papers in source corpus 40 papers cited in narrative 40 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

SLC1A2 (EAAT2/GLT-1) is the principal Na⁺-dependent high-affinity glutamate transporter in the mammalian forebrain, responsible for the bulk of synaptic glutamate clearance by astrocytes and also present in neuronal presynaptic terminals where it supplies glutamate to synaptic mitochondria for TCA cycle metabolism and protects against NMDA receptor–mediated excitotoxicity (PMID:10098717, PMID:12558972, PMID:30926746, PMID:35035352). Transport is electrogenic with an uncoupled anion conductance and voltage-dependent Na⁺ binding, and the transporter functions as a trimer whose activity can be enhanced allosterically at the trimerization–transport domain interface (PMID:25274824, PMID:29140675). Cell-surface abundance is regulated by PKC-induced ubiquitination at C-terminal lysines (K517, K526) triggering clathrin-dependent endocytosis and lysosomal degradation, counterbalanced by UCH-L1–mediated deubiquitination that promotes recycling through Rab4-positive endosomes, while SUMO1 conjugation retains a fraction intracellularly (PMID:17919781, PMID:18805448, PMID:22593014, PMID:24753081). Transcription is activated by NF-κB (EGF-dependent), PPARγ, Pax6, REST/CBP-p300, Notch signaling from endothelia, and KBBP/hnRNP K from neuronal signals, repressed by TNF-α/N-myc, and epigenetically controlled by CpG island and CpG shore methylation together with histone modifications (H3K27me3), with functional consequences for cortical spreading depression susceptibility, synaptic plasticity, brain development, and addiction-related glutamate homeostasis (PMID:15660126, PMID:17213861, PMID:26485579, PMID:34756885, PMID:28771710, PMID:19323997, PMID:17311293, PMID:22593010, PMID:32585762, PMID:19651762, PMID:16880397, PMID:24612076).

Mechanistic history

Synthesis pass · year-by-year structured walk · 22 steps
  1. 1998 High

    Establishing that EAAT2/GLT-1 is a Na⁺-dependent electrogenic glutamate transporter with ligand-gated channel properties resolved the biophysical basis of forebrain glutamate clearance.

    Evidence Expression cloning and electrophysiology in heterologous systems with subtype-specific pharmacology

    PMID:10098717

    Open questions at the time
    • stoichiometry of coupled ion movements not fully defined in this study
    • uncoupled anion conductance mechanism not resolved
  2. 1998 High

    Demonstrating neuronal expression of GLT-1 (previously considered glia-specific) and identification of a neuron-enriched splice variant overturned the assumption that EAAT2 functions exclusively in astrocytes.

    Evidence Immunocytochemistry, single-cell mRNA amplification, dihydrokainate-sensitive currents in hippocampal microcultures, cDNA cloning of GLT1v variant

    PMID:11784699 PMID:9614226 PMID:9761452

    Open questions at the time
    • relative contribution of neuronal vs. astrocytic EAAT2 to total clearance not quantified
    • functional significance of vesicular localization of GLT1v not determined
  3. 2003 High

    Showing that EAAT2 is the predominant nerve terminal glutamate transporter, with no detectable non-EAAT2 uptake in synaptosomes, established its presynaptic monopoly.

    Evidence Synaptosomal Western blotting, [³H]-glutamate uptake with dihydrokainate inhibition

    PMID:12558972

    Open questions at the time
    • role of presynaptic EAAT2 in glutamate recycling vs. metabolic supply not yet distinguished
  4. 2005 High

    Dissecting NF-κB as both an activator (EGF-dependent) and repressor (TNF-α/N-myc–dependent) of EAAT2 transcription revealed how opposing inflammatory and growth factor signals converge on the same promoter.

    Evidence Promoter mutagenesis, ChIP, IKKβ/p65 and N-myc overexpression in reporter assays

    PMID:15660126

    Open questions at the time
    • cell-type specificity of EGF vs. TNF-α pathways in vivo not established
    • chromatin context of NF-κB binding not examined
  5. 2006 High

    Identifying PPARγ response elements in the GLT-1 promoter and showing functional PPARγ-dependent upregulation linked metabolic/ischemic signaling to transporter transcription.

    Evidence Reporter assay, PPARγ agonist/antagonist, glutamate uptake, oxygen-glucose deprivation model

    PMID:17213861

    Open questions at the time
    • whether PPARγ directly binds all six PPREs in vivo not confirmed by ChIP
  6. 2006 High

    GLAST/GLT-1 double knockout mice revealed that glutamate transporter activity is essential for brain development, including cortical organization and neural stem cell proliferation.

    Evidence Double-knockout histology and immunohistochemistry of cortical, hippocampal, and olfactory bulb development

    PMID:16880397

    Open questions at the time
    • individual contribution of GLT-1 vs. GLAST to each developmental phenotype not separated
    • extracellular glutamate levels not directly measured in embryonic brain
  7. 2007 High

    Demonstrating that CpG island methylation silences EAAT2 transcription established an epigenetic mechanism for transporter loss in glioma and potentially in neurodegeneration.

    Evidence Bisulfite sequencing of glioma vs. brain tissue, in vitro methylation reporter assay, EMSA

    PMID:17311293

    Open questions at the time
    • writers/erasers responsible for methylation changes not identified
    • relevance to ALS later shown to be minimal (PMID:19672971)
  8. 2007 High

    Mapping the PKC-triggered endocytic route as clathrin/dynamin-dependent (not caveolae or Arf6) with lysosomal degradation via Rab7 defined the internalization pathway for EAAT2.

    Evidence Surface biotinylation with dominant-negative dynamin, clathrin, Rab7, caveolin, Eps15, Arf6 constructs; lysosomal inhibitors

    PMID:17919781

    Open questions at the time
    • adaptor proteins linking ubiquitinated GLT-1 to clathrin machinery not identified
  9. 2008 High

    Identifying redundant C-terminal lysines as the PKC-dependent ubiquitination sites on GLT-1 provided the molecular link between kinase signaling and transporter internalization.

    Evidence Site-directed mutagenesis of all 11 cytoplasmic lysines, ubiquitin incorporation in C6 glioma and cortical cultures

    PMID:18805448

    Open questions at the time
    • E3 ubiquitin ligase responsible not identified in this study
  10. 2009 High

    Discovery that neuronal signals maintain astrocytic GLT-1 through KBBP/hnRNP K binding the GLT-1 promoter explained why denervation and motor neuron disease reduce astroglial glutamate transport.

    Evidence Promoter binding assays, neuron-astrocyte co-cultures, in vivo denervation, ALS mouse model

    PMID:19323997

    Open questions at the time
    • signal from neurons to astrocytes that induces KBBP not molecularly defined
  11. 2009 High

    Showing that upregulated GLT-1 at mossy fiber terminals impairs mGluR-dependent LTD and LTP demonstrated that presynaptic EAAT2 levels gate synaptic plasticity by controlling perisynaptic glutamate concentration.

    Evidence Ceftriaxone-induced GLT-1 upregulation, electrophysiology, dihydrokainate rescue, immunogold EM, GLT-1 KO controls

    PMID:19651762

    Open questions at the time
    • whether endogenous regulation of presynaptic GLT-1 modulates plasticity under physiological conditions not tested
  12. 2012 High

    Defining constitutive GLT-1 recycling through Rab4-positive endosomes, with K517/K526 ubiquitination driving internalization and UCH-L1 deubiquitination promoting recycling, completed the bidirectional trafficking model.

    Evidence Clathrin inhibitors, Rab dominant-negatives, site-directed K517R/K526R mutagenesis, UCH-L1 inhibitor in astrocytes

    PMID:22593014

    Open questions at the time
    • whether K517/K526 are the same sites modified by PKC or represent constitutive-only sites not fully resolved
  13. 2012 High

    Identification of Pax6 binding a distal enhancer ~8 kb upstream of GLT-1 and the CpG shore as a methylation-sensitive enhancer element expanded the cis-regulatory landscape controlling region-specific expression.

    Evidence ChIP, EMSA, lentiviral overexpression/shRNA knockdown; targeted in vitro methylation + H3K27me3 ChIP in cortex vs. cerebellum astrocytes

    PMID:22593010 PMID:26485579

    Open questions at the time
    • combinatorial interactions among Pax6, NF-κB, PPARγ, and epigenetic marks not tested
  14. 2014 High

    Reconstituted liposome studies resolved that EAAT2 heteroexchange and net uptake have comparable rates, and that voltage-dependent third Na⁺ binding underlies voltage sensitivity, clarifying the transporter's biophysical mechanism.

    Evidence Purified EAAT2 in liposomes, voltage manipulation, anion substitution, computational modeling

    PMID:25274824

    Open questions at the time
    • structural basis for voltage-dependent Na⁺ binding site not determined
  15. 2014 High

    Discovery that SUMO1 conjugation retains EAAT2 intracellularly while non-sumoylated EAAT2 resides at the plasma membrane added a second post-translational code (beyond ubiquitin) controlling surface availability.

    Evidence SUMO1-EAAT2 co-IP, subcellular fractionation, desumoylation promotion with functional uptake in primary astrocytes and SOD1-G93A mice

    PMID:24753081

    Open questions at the time
    • SUMO E3 ligase and desumoylating enzyme acting on EAAT2 not identified
    • interplay between sumoylation and ubiquitination not examined
  16. 2017 High

    Identifying an allosteric activation site at the trimerization–transport domain interface showed that EAAT2 translocation rate can be pharmacologically enhanced without altering substrate binding.

    Evidence Virtual screening, mutagenesis of interface residues, transport assays in heterologous cells

    PMID:29140675

    Open questions at the time
    • structural model at atomic resolution not available
    • in vivo efficacy of allosteric modulators not tested
  17. 2017 High

    Demonstrating contact-dependent Notch signaling from brain endothelia to astrocytes as a GLT-1 inducer revealed a vascular–glial axis for transporter regulation.

    Evidence Astrocyte-endothelial co-culture with transwell, γ-secretase inhibitor DAPT, RBPJκ shRNA, GLT-1 eGFP reporter

    PMID:28771710

    Open questions at the time
    • specific Notch ligand on endothelia not identified
    • in vivo validation with endothelial-specific Notch ligand deletion not performed
  18. 2019 High

    Conditional neuronal EAAT2 deletion with isotope tracing proved that presynaptic EAAT2 supplies glutamate to synaptic mitochondria for TCA metabolism and that its loss remodels mitochondrial bioenergetics.

    Evidence Synapsin1-Cre conditional KO, U-¹³C-glucose and U-¹³C-glutamate tracing, synaptosomal uptake, mitochondrial ATP/O₂ assays, EM for cristae density

    PMID:30926746

    Open questions at the time
    • whether neuronal EAAT2 metabolic role extends to inhibitory terminals not tested
  19. 2019 High

    Cell-type-specific conditional deletions separated astrocytic EAAT2 (early memory deficits, inflammatory/synaptic transcriptomic overlap with AD) from neuronal EAAT2 (late-onset spatial deficit, kynurenine pathway dysregulation), demonstrating non-redundant in vivo functions.

    Evidence Astrocytic vs. neuronal conditional KO mice, Morris water maze, novel object recognition, transcriptomics

    PMID:31591195

    Open questions at the time
    • direct causal link between kynurenine pathway changes and neuronal EAAT2 loss not established
  20. 2020 High

    Conditional glial GLT-1 knockout increased cortical spreading depression frequency and velocity with accelerated extracellular glutamate accumulation, establishing GLT-1 as the critical transporter gating CSD susceptibility.

    Evidence Conditional KO, electrophysiological CSD recording, enzyme-based glutamate biosensor, comparison with EAAC1 and GLAST KOs

    PMID:32585762

    Open questions at the time
    • whether GLT-1 loss affects CSD through glutamate clearance alone or also via uncoupled conductance not distinguished
  21. 2021 High

    Demonstrating that neuronal GLT-1 loss causes NMDA receptor–dependent excitotoxic synaptic failure, rescuable by MK-801 or glutamate scavenging, linked the metabolic and neuroprotective roles of presynaptic EAAT2.

    Evidence Conditional neuronal KO, hippocampal field recordings, extracellular FRET glutamate sensor, MK-801 rescue, EM

    PMID:35035352

    Open questions at the time
    • relative contribution of impaired mitochondrial metabolism vs. extracellular glutamate buildup to excitotoxicity not fully separated
  22. 2021 High

    Identification of REST/CBP-p300 as a transcriptional activator of EAAT2 in astrocytes, with neuroprotective consequences against manganese-induced excitotoxicity, added an epigenetic co-activator axis to the regulatory network.

    Evidence REST overexpression/knockdown, ChIP for CBP/p300 at EAAT2 promoter, glutamate uptake, astrocyte-neuron co-culture death assay

    PMID:34756885

    Open questions at the time
    • whether REST acts through the same or distinct cis-elements as NF-κB and Pax6 not mapped

Open questions

Synthesis pass · forward-looking unresolved questions
  • The E3 ubiquitin ligase targeting EAAT2 for internalization, the SUMO E3 ligase/desumoylase pair controlling intracellular retention, the structural basis for allosteric activation and voltage-dependent Na⁺ binding, and the molecular identity of the neuronal signal that activates KBBP in astrocytes remain unidentified.
  • E3 ligase for EAAT2 ubiquitination unknown
  • SUMO machinery acting on EAAT2 unidentified
  • high-resolution structure of EAAT2 with allosteric modulator not solved
  • neuronal signal upstream of KBBP/hnRNP K not molecularly defined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005215 transporter activity 4 GO:0140657 ATP-dependent activity 2
Localization
GO:0005886 plasma membrane 4 GO:0005768 endosome 2 GO:0031410 cytoplasmic vesicle 2 GO:0005829 cytosol 1
Pathway
R-HSA-74160 Gene expression (Transcription) 7 R-HSA-112316 Neuronal System 5 R-HSA-162582 Signal Transduction 5 R-HSA-382551 Transport of small molecules 4 R-HSA-392499 Metabolism of proteins 3 R-HSA-5653656 Vesicle-mediated transport 3 R-HSA-4839726 Chromatin organization 2 R-HSA-1266738 Developmental Biology 1

Evidence

Reading pass · 40 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1998 GLT-1/EAAT2 functions as a Na+-dependent high-affinity glutamate transporter responsible for the bulk of forebrain glutamate uptake, and biophysical studies of cloned transporters revealed that some subtypes also function as ligand-gated ion channels. Expression systems, biophysical/electrophysiological recordings, subtype-specific antibodies Neurochemistry international High 10098717
1998 GLT-1/EAAT2 protein is expressed in neurons (hippocampal microcultures), primarily in dendrites of excitatory neurons, and neuronal GLT-1 can participate in clearance of synaptically released glutamate as shown by dihydrokainate-sensitive transport currents and prolonged autaptic currents after glial inactivation. Immunocytochemistry with N- and C-terminal antibodies, whole-cell electrophysiology, pharmacological blockade with dihydrokainate The Journal of neuroscience High 9614226
1998 GLT-1 (EAAT2) mRNA and protein are expressed in cultured hippocampal neurons (not restricted to glia), demonstrated by single-cell mRNA amplification and immunocytochemistry. Single-cell mRNA amplification, immunocytochemistry with subtype-specific antibodies Neurochemistry international Medium 9761452
1998 A splice variant of GLT1/EAAT2 (GLT1v) generated by alternative splicing at the 3'-end is preferentially expressed in neurons (CNS and PNS) rather than astrocytes, and immunolabeling shows a cytoplasmic/granular localization suggesting vesicle membrane association. cDNA cloning, RT-PCR, Northern blot, in situ hybridization, immunocytochemistry Neuroscience High 11784699
1998 Alternative splicing of human EAAT2 produces two novel transcripts (EAAT2-C1, lacking exon 8; EAAT2-C2, with internal splice sites in exons 5 and 6) resulting in deletions of 45 and 107 amino acids in the C-terminal and central protein regions, respectively. RT-PCR cloning, sequence analysis Neuroscience letters Medium 9502218
2003 EAAT2 (GLT-1) is the predominant nerve terminal glutamate transporter in adult rodent CNS; Western blotting detected EAAT2 in synaptosomes, and pharmacological inhibition with dihydrokainate failed to unmask any non-EAAT2 uptake sites in nerve terminals. Western blotting of synaptosomes and glial plasmalemmal vesicles, [3H]D-aspartate and [3H]L-glutamate uptake assays, pharmacological inhibition Journal of neurochemistry High 12558972
2005 NF-κB positively regulates EAAT2 transcription (EGF-dependent activation), while TNFα mediates repression of EAAT2 through a distinct NF-κB pathway requiring recruitment of N-myc to N-myc binding sites in the EAAT2 promoter; EGF activates NF-κB independently of IκB signaling. Reporter gene assays, ChIP, promoter mutagenesis, IKKβ/p65 overexpression, N-myc overexpression The EMBO journal High 15660126
2007 DNA methylation of the EAAT2 promoter CpG island silences EAAT2 expression in glial cells; bisulfite analysis showed dense methylation in EAAT2-negative glioma lines vs. hypomethylation in EAAT2-positive brain tissue; in vitro methylation reduced promoter activity and altered nuclear factor binding. Bisulfite sequencing, EMSA, reporter gene assay, DNA methyltransferase inhibition Glia High 17311293
2007 PKC activation causes clathrin-dependent endocytosis and subsequent lysosomal degradation of GLT-1; dominant-negative dynamin1, clathrin heavy chain, and Rab7 constructs blocked these effects, while cholesterol depletion, caveolin-1 DN, Eps15 DN, and Arf6 DN had no effect. Surface biotinylation, dominant-negative constructs (dynamin, clathrin, Rab7, caveolin, Eps15, Arf6), lysosomal inhibitors (chloroquine, ammonium chloride) Neurochemistry international High 17919781
2008 PKC activation induces ubiquitination of GLT-1 at redundant lysine residues in the carboxyl terminus (specifically C7K-R region), which is required for PKC-dependent internalization and degradation; mutation of all 11 cytoplasmic lysines abolished ubiquitination. Immunoprecipitation, site-directed mutagenesis of lysine residues, ubiquitin incorporation assay in C6 glioma and primary cortical cultures Neurochemistry international High 18805448
2009 Presynaptic terminals regulate astroglial GLT-1/EAAT2 expression via kappa B-motif binding phosphoprotein (KBBP/hnRNP K), which binds the GLT-1 promoter and is required for transcriptional activation; denervation reduces KBBP expression and causes astroglial transporter dysfunction. Promoter binding assays, neuron-astrocyte co-culture, in vivo denervation models (corticospinal tract transection, ricin-induced motor neuron death), ALS mouse model Neuron High 19323997
2009 PIKfyve (phosphatidylinositol-3-phosphate-5-kinase) enhances EAAT2-mediated glutamate transport current and increases EAAT2 protein abundance at the cell membrane; this effect depends on SGK1 phosphorylation of PIKfyve at S318. Xenopus oocyte expression, electrophysiology (glutamate-induced inward currents), confocal microscopy, mutagenesis (S318A PIKfyve), kinase-dead SGK1 construct Cellular physiology and biochemistry High 19910676
2010 DNA demethylation of selective CpG sites in the GLT-1 promoter correlates with increased GLT-1 mRNA in astrocytes in response to neuronal stimulation; hypermethylation at selective CpG sites represses GLT-1 promoter activation, but this mechanism does not account for EAAT2 dysregulation in ALS. Bisulfite sequencing of FACS-isolated astrocytes from BAC GLT-1 eGFP mice, in vitro and in vivo neuronal stimulation paradigms, postmortem ALS motor cortex analysis Glia High 19672971
2011 GLT-1 co-compartmentalizes with mitochondria and glycolytic enzymes (including hexokinase-1) in fine astrocytic processes; immunoaffinity isolation identified these interacting proteins by LC-MS/MS, and simultaneous inhibition of both glycolysis and oxidative phosphorylation (but not either alone) significantly reduces glutamate transport. GLT-1 immunoaffinity isolation from rat cortex, LC-MS/MS proteomics, double-label immunofluorescence, biolistic transfection in hippocampal slices, Monte Carlo simulation, acute metabolic inhibition assays The Journal of neuroscience High 22171032
2012 Constitutive GLT-1 internalization occurs via clathrin-dependent endocytosis into EEA1/Rab4-positive recycling endosomes (not Rab11 or Rab7 compartments); ubiquitination (at lysines 517 and 526) drives internalization, and deubiquitination by UCH-L1 promotes recycling to the plasma membrane. Clathrin inhibitors, dominant-negative Rab constructs, E1 ubiquitin enzyme inhibitor, site-directed mutagenesis (K517, K526), UCH-L1 inhibitor (LDN-57444), endosomal marker co-localization in heterologous system and primary astrocytes Glia High 22593014
2012 The transcription factor Pax6 is expressed in astrocytes and contributes to neuron-induced GLT-1 expression by binding to a conserved distal enhancer element ~8 kb upstream of the GLT-1 translation start site. Lentiviral Pax6 overexpression in BAC GLT-1 eGFP reporter astrocytes, shRNA knockdown, EMSA, ChIP, GLT-1 protein and uptake assays Journal of neurochemistry High 26485579
2013 Astroglial FMRP positively regulates mGluR5 protein translation in astrocytes (FMRP associates with mGluR5 mRNA); loss of astroglial FMRP reduces mGluR5 protein and Ca2+ responses, which in turn attenuates neuron-dependent GLT-1 expression and glutamate uptake in fmr1-/- mice. Mismatched astrocyte-neuron co-cultures, FMRP immunoprecipitation + qRT-PCR, Western blot, Ca2+ imaging, dihydrokainate-sensitive GLT-1 inhibition in cortical slices Human molecular genetics High 23396537
2014 EAAT2 undergoes constitutive sumoylation (SUMO1 conjugation) in astrocytes in vitro and in vivo; sumoylated EAAT2 localizes to intracellular compartments while non-sumoylated EAAT2 resides on the plasma membrane; promoting desumoylation increases EAAT2-mediated glutamate uptake. Immunoprecipitation for SUMO1-EAAT2, immunofluorescence, subcellular fractionation, desumoylation promotion assay with functional glutamate uptake measurement in primary astrocytes and SOD1-G93A mouse Glia High 24753081
2014 EAAT2 heteroexchange and net uptake have comparable rates; net uptake is sensitive to membrane potential and stimulated by external permeable anions (uncoupled anion conductance); a sodium leak is also present in EAAT2; the voltage sensitivity of exchange is caused by voltage-dependent third Na+ binding. Reconstituted liposome system with rat and mouse EAAT2 protein, electrophysiological voltage manipulation, anion substitution, computational modeling The Journal of neuroscience High 25274824
2014 Small-molecule LDN/OSU-0212320 activates EAAT2 translation through PKC activation and subsequent Y-box-binding protein 1 (YB-1) phosphorylation, which drives EAAT2 translational activation. Cell-based translational activation assays, PKC pathway inhibitors, YB-1 phosphorylation analysis, in vivo pharmacokinetics and ALS/epilepsy mouse models The Journal of clinical investigation High 24569372
2017 Novel positive allosteric modulators of EAAT2 interact with residues at the interface between the trimerization domain and the substrate-binding transport domain; mutagenesis of these residues abolished activator effects; activators enhance glutamate translocation rate without affecting substrate binding, confirming an allosteric mechanism. Virtual screening, in vitro transport assay in heterologous expression system, site-directed mutagenesis at trimerization/transport domain interface, selectivity assays ACS chemical neuroscience High 29140675
2017 Brain endothelial cells induce astrocytic GLT-1 expression through a contact-dependent, Notch-dependent mechanism; γ-secretase inhibition blocks endothelia-induced Notch intracellular domain nuclear accumulation and GLT-1 upregulation; shRNA against RBPJκ (Notch effector) reduces endothelial induction of GLT-1. Astrocyte-endothelial co-culture with transwells (contact dependence), γ-secretase inhibitor (DAPT), shRNA against RBPJκ, BAC GLT-1 eGFP reporter mice, GLT-1 protein and glutamate uptake assays Journal of neurochemistry High 28771710
2018 Neuronal EAAT2 in axon terminals is present throughout multiple forebrain regions (not just hippocampus); conditional neuronal deletion (synapsin1-Cre) disproportionately reduces glutamate accumulation in homogenates relative to protein loss, and increases 13C-labeling of glutamine and GABA suggesting neuronal EAAT2 partially short-circuits the glutamate-glutamine cycle. Conditional KO (synapsin1-Cre × EAAT2-flox), Western blot, glutamate accumulation in tissue homogenates, U-13C-glucose isotope tracing, Ai9 Cre-reporter Neurochemistry international High 29530756
2019 Neuronal GLT-1 (synGLT-1 KO) is required to supply glutamate to synaptic mitochondria for TCA cycle metabolism; its loss reduces aspartate content, diminishes [U-13C]-glutamate-derived TCA labeling, decreases pyruvate recycling, increases mitochondrial ATP production efficiency, and increases mitochondrial cristae density in axon terminals. Conditional KO (synapsin1-Cre), synaptosomal uptake, electron microscopy immunocytochemistry, 13C-isotope tracing, isolated mitochondria ATP/oxygen consumption assays The Journal of neuroscience High 30926746
2019 Astrocytic conditional EAAT2 deletion causes early deficits in short-term and spatial long-term memory with transcriptomic signatures overlapping human AD and aging (inflammatory/synaptic pathways); neuronal EAAT2 deletion causes late-onset spatial memory deficit with kynurenine pathway dysregulation—demonstrating cell-type-specific roles. Conditional KO mice (astrocytic vs. neuronal EAAT2 deletion), behavioral testing (Morris water maze, novel object recognition), transcriptomics Proceedings of the National Academy of Sciences High 31591195
2021 Loss of neuronal GLT-1 in presynaptic terminals causes excitotoxic failure of synaptic transmission in CA1; NMDA receptor blockade (MK801) or glutamate scavenging prevents fEPSP failure, indicating that neuronal GLT-1 protects against NMDA receptor-mediated excitotoxicity; metabolic perturbations (reduced glutamate utilization by synaptic mitochondria) contribute to vulnerability. Conditional neuronal GLT-1 KO, field potential recordings in hippocampal slices, extracellular FRET-based glutamate sensor, MK801 pharmacology, electron microscopy for mitochondrial cristae density Frontiers in cellular neuroscience High 35035352
2011 CD44-SLC1A2 gene fusion in gastric cancer (caused by paracentric chromosomal inversion) places SLC1A2 under CD44 regulatory elements, driving its overexpression; silencing CD44-SLC1A2 reduces intracellular glutamate, proliferation, invasion, and anchorage-independent growth, while overexpression promotes these pro-oncogenic traits. Genomic breakpoint analysis, RT-PCR for fusion transcript, siRNA knockdown, overexpression in gastric cell lines, proliferation/invasion/anchorage-independent growth assays, intracellular glutamate measurement Science translational medicine High 21471434
2006 PPARγ is a transcriptional regulator of GLT-1/EAAT2; rosiglitazone (PPARγ agonist) increases GLT-1 mRNA and protein and [3H]glutamate uptake; six PPAR response elements (PPREs) were identified in the GLT-1 promoter; rosiglitazone increased GLT-1 promoter activity 4-fold; PPARγ antagonist blocks ischemic preconditioning-induced GLT-1 upregulation. Reporter gene assay, [3H]glutamate uptake, Western blot, RT-PCR, PPARγ antagonist/agonist pharmacology, in vitro oxygen-glucose deprivation model Journal of cerebral blood flow and metabolism High 17213861
2012 GPR30 activation (by G1 agonist) increases GLT-1 protein and mRNA in astrocytes through MAPK and PI3K signaling, TGF-α receptor transactivation, PKA, and NF-κB (both p50 and p65 subunits bind the GLT-1 promoter); GPR30 silencing reduces GLT-1 and TGF-α expression. GPR30 siRNA knockdown, G1 agonist treatment, MAPK/PI3K inhibitors, PKA inhibitor, NF-κB inhibitor, CREB and NF-κB ChIP on GLT-1 promoter, glutamate uptake assay in rat primary astrocytes The Journal of biological chemistry High 22645130
2011 Equilibrative nucleoside transporter 1 (ENT1) regulates EAAT2 expression and function in astrocytes; ENT1 antagonist or siRNA reduces EAAT2 mRNA and glutamate uptake; ENT1 overexpression upregulates EAAT2; ENT1 knockdown inhibits ethanol-induced EAAT2 upregulation. ENT1-specific antagonist, siRNA knockdown, overexpression, qRT-PCR, glutamate uptake assay in cultured astrocytes Alcoholism, clinical and experimental research Medium 20374202
2006 Glucocorticoid receptor activation by corticosterone inhibits microglial GLT-1 expression and glutamate uptake, likely through suppression of TNF-α release; mifepristone (glucocorticoid receptor antagonist) blocks this effect; exogenous TNF-α counteracts corticosterone's inhibitory effect on GLT-1. Primary microglial cultures, LPS stimulation, corticosterone dose-response, mifepristone antagonism, TNF-α measurement, GLT-1 expression (Western blot/immunostaining), glutamate uptake assay Neuroscience Medium 16473474
2020 Glial GLT-1 determines susceptibility to cortical spreading depression (CSD); conditional GLT-1 KO mice show enhanced CSD frequency and velocity, more rapid extracellular glutamate accumulation during early CSD phase (measured by enzyme-based biosensor), while EAAC1 and GLAST germline KOs show no such effect. Conditional GLT-1 KO mice, electrophysiological CSD recording, hemodynamic imaging, enzyme-based extracellular glutamate biosensor Glia High 32585762
2021 Astrocytic REST transcription factor positively regulates EAAT2 expression by recruiting the epigenetic co-activator CBP/p300 to REST binding sites in the EAAT2 promoter; REST overexpression in astrocytes attenuates Mn-induced reduction of EAAT2 and prevents excitotoxic dopaminergic neuronal death in co-culture. REST overexpression/knockdown in astrocytes, ChIP for CBP/p300 at EAAT2 promoter, glutamate uptake assay, astrocyte-neuron co-culture dopaminergic death assay The Journal of biological chemistry High 34756885
2012 The CpG island shore of the GLT-1 gene acts as a methylation-sensitive enhancer; in vitro methylation of shore CpG sites abolishes dexamethasone-stimulated transcriptional activity; the shore region shows higher methylation and repressive histone marks (H3K27me3) in cerebellum (low GLT-1) vs. cortex (high GLT-1) astrocytes. Reporter gene assay, targeted in vitro CpG methylation, ChIP for H3K27me3 and H4ac, bisulfite sequencing, HDAC inhibitor treatment in region-specific astrocytes Glia High 22593010
2016 The laforin/malin complex (mutated in Lafora disease) slows endocytic recycling of GLT-1; overexpression of laforin and malin causes accumulation of GLT-1 at the plasma membrane and reduces GLT-1 ubiquitination; primary astrocytes from Lafora disease mice have reduced GLT-1 at plasma membrane and reduced glutamate transport capacity. Laforin/malin overexpression in cellular models, surface GLT-1 quantification, ubiquitination assays, glutamate transport assay in primary Lafora disease mouse astrocytes Biochimica et biophysica acta Medium 26976331
2009 Up-regulation of GLT-1 by ceftriaxone severely impairs mGluR-dependent LTD at mossy fiber-CA3 synapses and reduces LTP at the same synapses; postembedding immunogold shows increased GLT-1a density at mossy fiber terminals near and within active zones, revealing that presynaptic GLT-1 prevents activation of presynaptic receptors needed for plasticity. Ceftriaxone treatment, field potential recordings, dihydrokainate rescue, gamma-DGG assay for synaptic glutamate transient, postembedding immunogold electron microscopy, GLT-1 KO mice as specificity controls The Journal of physiology High 19651762
2006 GLAST/GLT-1 double knockout mice show multiple brain developmental defects (cortical, hippocampal, olfactory bulb disorganization), impaired neural stem cell proliferation, radial migration, neuronal differentiation, and survival of subplate neurons, demonstrating that GLAST and GLT-1 are essential for brain development through regulation of extracellular glutamate. GLAST/GLT-1 double knockout mouse, histology, immunohistochemistry, analysis of cortical development Proceedings of the National Academy of Sciences High 16880397
2013 Proteome analysis of pancreatic islets detected that EAAT2 levels are too low to support any proposed glutamate transport functions in islets; conditional pancreatic EAAT2 deletion did not affect survival, growth, glucose tolerance, or β-cell number, ruling out a role for EAAT2 in insulin secretion. Conditional pancreatic EAAT2 KO (RIP-Cre, IPF1-Cre), LC-MS/MS proteomics of islet proteins (>7000 proteins detected), TaqMan RT-PCR, immunoblotting, immunocytochemistry, glucose tolerance testing The Journal of biological chemistry High 24280215
2014 Restoring GLT-1 expression (but not xCT/cystine-glutamate exchange) in nucleus accumbens is the key mechanism by which chronic N-acetylcysteine (NAC) inhibits cue-induced cocaine reinstatement; suppressing NAC-induced GLT-1 restoration with vivo-morpholino antisense augmented reinstatement via increased extracellular glutamate activating mGluR5. Rat self-administration/extinction/reinstatement model, vivo-morpholino antisense oligomers targeting GLT-1 or xCT, mGluR5 blockade, intra-accumbal microinjection Addiction biology High 24612076
2004 Adenoviral overexpression of GLT-1 specifically in the locus coeruleus inhibits naloxone-precipitated morphine withdrawal signs, demonstrating that GLT-1 in the locus coeruleus plays an inhibitory role in morphine physical dependence. Recombinant adenovirus-mediated GLT-1 gene transfer into bilateral locus coeruleus, morphine pellet implantation, naloxone-precipitated withdrawal scoring in rats The European journal of neuroscience Medium 14750980

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2019 The role of astrocytic glutamate transporters GLT-1 and GLAST in neurological disorders: Potential targets for neurotherapeutics. Neuropharmacology 347 30851309
2011 Role of excitatory amino acid transporter-2 (EAAT2) and glutamate in neurodegeneration: opportunities for developing novel therapeutics. Journal of cellular physiology 323 21792905
2011 Molecular comparison of GLT1+ and ALDH1L1+ astrocytes in vivo in astroglial reporter mice. Glia 209 21046559
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