Home LiteratureArticle Details
PMID: 14762132 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The glutamate transporter GLT1a is expressed in excitatory axon terminals of mature hippocampal neurons.

Chen W, Mahadomrongkul V, Berger UV, Bassan M, DeSilva T, Tanaka K, Irwin N, Aoki C, Rosenberg PA

Abstract

GLT1 is the major glutamate transporter of the brain and has been thought to be expressed exclusively in astrocytes. Although excitatory axon terminals take up glutamate, the transporter responsible has not been identified. GLT1 is expressed in at least two forms varying in the C termini, GLT1a and GLT1b. GLT1 mRNA has been demonstrated in neurons, without associated protein. Recently, evidence has been presented, using specific C terminus-directed antibodies, that GLT1b protein is expressed in neurons in vivo. These data suggested that the GLT1 mRNA detected in neurons encodes GLT1b and also that GLT1b might be the elusive presynaptic transporter. To test these hypotheses, we used variant-specific probes directed to the 3'-untranslated regions for GLT1a and GLT1b to perform in situ hybridization in the hippocampus. Contrary to expectation, GLT1a mRNA was the more abundant form. To investigate further the expression of GLT1 in neurons in the hippocampus, antibodies raised against the C terminus of GLT1a and against the N terminus of GLT1, found to be specific by testing in GLT1 knock-out mice, were used for light microscopic and EM-ICC. GLT1a protein was detected in neurons, in 14-29% of axons in the hippocampus, depending on the region. Many of the labeled axons formed axo-spinous, asymmetric, and, thus, excitatory synapses. Labeling also occurred in some spines and dendrites. The antibody against the N terminus of GLT1 also produced labeling of neuronal processes. Thus, the originally cloned form of GLT1, GLT1a, is expressed as protein in neurons in the mature hippocampus and may contribute significantly to glutamate uptake into excitatory terminals.

MeSH Terms
Animals Antibody Specificity Brain Chemistry Excitatory Amino Acid Transporter 2/biosynthesis,genetics Heterozygote Hippocampus/cytology,metabolism,ultrastructure Homozygote Immunohistochemistry In Situ Hybridization/methods Mice Mice, Knockout Neurons/metabolism,ultrastructure Presynaptic Terminals/metabolism,ultrastructure Protein Isoforms/biosynthesis,genetics Protein Transport/physiology RNA, Messenger/analysis,biosynthesis Rats Rats, Sprague-Dawley
Chemicals
Excitatory Amino Acid Transporter 2 Protein Isoforms RNA, Messenger
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Chen Weizhi
Department of Neurology, Children's Hospital, Boston, Massachusetts 02115, USA.
Mahadomrongkul Veeravan
Berger Urs V
Bassan Merav
DeSilva Tara
Tanaka Kohichi
Irwin Nina
Aoki Chiye
Rosenberg Paul A
References (73)
73 references, click to expand
  1. Metabolic trafficking between neurons and astrocytes: the glutamate/glutamine cycle revisited.
    Dev Neurosci. 1995;17(4):203-11 PMID: 8575339
  2. Expression of the glutamate transporter GLT1 in neural cells of the rat central nervous system: non-radioactive in situ hybridization and comparative immunocytochemistry.
    Neuroscience. 1996 Apr;71(4):989-1004 PMID: 8684627
  3. Differential expression of three glutamate transporter subtypes in the rat retina.
    Cell Tissue Res. 1996 Dec;286(3):325-36 PMID: 8929335
  4. Selective excitatory amino acid uptake in glutamatergic nerve terminals and in glia in the rat striatum: quantitative electron microscopic immunocytochemistry of exogenous (D)-aspartate and endogenous glutamate and GABA.
    Eur J Neurosci. 1996 Apr;8(4):758-65 PMID: 9081627
  5. Extrasynaptic glutamate spillover in the hippocampus: dependence on temperature and the role of active glutamate uptake.
    Neuron. 1997 Feb;18(2):281-93 PMID: 9052798
  6. Differential distribution of the glutamate transporters GLT1 and rEAAC1 in rat cerebral cortex and thalamus: an in situ hybridization analysis.
    Anat Embryol (Berl). 1997 Apr;195(4):317-26 PMID: 9108197
  7. Epilepsy and exacerbation of brain injury in mice lacking the glutamate transporter GLT-1.
    Science. 1997 Jun 13;276(5319):1699-702 PMID: 9180080
  8. Hypoxia-ischemia causes abnormalities in glutamate transporters and death of astroglia and neurons in newborn striatum.
    Ann Neurol. 1997 Sep;42(3):335-48 PMID: 9307255
  9. Tissue specific variants of glutamate transporter GLT-1.
    FEBS Lett. 1997 Oct 27;416(3):312-6 PMID: 9373176
  10. Metabolic compartmentation of glutamate and glutamine: morphological evidence obtained by quantitative immunocytochemistry in rat cerebellum.
    Neuroscience. 1992;46(3):519-34 PMID: 1347649
  11. A monoclonal antibody against a Na(+)-L-glutamate cotransporter from rat brain.
    J Biol Chem. 1992 Nov 15;267(32):23275-81 PMID: 1429674
  12. Structure, expression, and functional analysis of a Na(+)-dependent glutamate/aspartate transporter from rat brain.
    Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10955-9 PMID: 1279699
  13. Cloning and expression of a rat brain L-glutamate transporter.
    Nature. 1992 Dec 3;360(6403):464-7 PMID: 1448170
  14. An [Na+ + K+]coupled L-glutamate transporter purified from rat brain is located in glial cell processes.
    Neuroscience. 1992 Nov;51(2):295-310 PMID: 1465194
  15. Subtypes of sodium-dependent high-affinity L-[3H]glutamate transport activity: pharmacologic specificity and regulation by sodium and potassium.
    J Neurochem. 1993 Jan;60(1):167-79 PMID: 8093259
  16. A monoclonal antibody raised against an [Na(+)+K+]coupled L-glutamate transporter purified from rat brain confirms glial cell localization.
    FEBS Lett. 1993 Feb 8;317(1-2):79-84 PMID: 7679083
  17. Ultrastructural immunocytochemical observations on the localization, metabolism and transport of glutamate in normal and ischemic brain tissue.
    Prog Brain Res. 1992;94:225-41 PMID: 1363142
  18. Metabolism of [U-13C]glutamate in astrocytes studied by 13C NMR spectroscopy: incorporation of more label into lactate than into glutamine demonstrates the importance of the tricarboxylic acid cycle.
    J Neurochem. 1993 Sep;61(3):1179-82 PMID: 8103082
  19. Adenosine triphosphate depletion reverses sodium-dependent, neuronal uptake of glutamate in rat hippocampal slices.
    J Neurosci. 1993 Oct;13(10):4429-44 PMID: 8105040
  20. Demonstration of glutamate/aspartate uptake activity in nerve endings by use of antibodies recognizing exogenous D-aspartate.
    Neuroscience. 1993 Nov;57(1):97-111 PMID: 7904057
  21. Localization of the glutamate transporter GLT-1 in rat and macaque monkey retinae.
    Neurosci Lett. 1994 Mar 14;169(1-2):137-40 PMID: 8047270
  22. Functional comparisons of three glutamate transporter subtypes cloned from human motor cortex.
    J Neurosci. 1994 Sep;14(9):5559-69 PMID: 7521911
  23. Localization of neuronal and glial glutamate transporters.
    Neuron. 1994 Sep;13(3):713-25 PMID: 7917301
  24. Differential expression of two glial glutamate transporters in the rat brain: an in situ hybridization study.
    Eur J Neurosci. 1994 Jun 1;6(6):936-42 PMID: 7952280
  25. Kinetics of 2-oxoglutarate uptake by synaptosomes from bovine and rat retina and cerebral cortex and regulation by glutamate and glutamine.
    Dev Neurosci. 1993;15(3-5):330-5 PMID: 7805586
  26. Astrocytes: glutamate producers for neurons.
    J Neurosci Res. 1999 Aug 15;57(4):417-28 PMID: 10440891
  27. Differentiation of substrate and nonsubstrate inhibitors of the high-affinity, sodium-dependent glutamate transporters.
    Mol Pharmacol. 1999 Dec;56(6):1095-104 PMID: 10570036
  28. Glutamate release in severe brain ischaemia is mainly by reversed uptake.
    Nature. 2000 Jan 20;403(6767):316-21 PMID: 10659851
  29. Neuronal pyruvate carboxylation supports formation of transmitter glutamate.
    J Neurosci. 2000 Feb 15;20(4):1342-7 PMID: 10662824
  30. Use of electron microscopy in the detection of adrenergic receptors.
    Methods Mol Biol. 2000;126:535-63 PMID: 10685434
  31. Cerebral metabolism of lactate in vivo: evidence for neuronal pyruvate carboxylation.
    J Cereb Blood Flow Metab. 2000 Feb;20(2):327-36 PMID: 10698070
  32. Differential synaptic localization of the glutamate transporter EAAC1 and glutamate receptor subunit GluR2 in the rat hippocampus.
    J Comp Neurol. 2000 Mar 13;418(3):255-69 PMID: 10701825
  33. Distribution of the glutamate transporters GLAST and GLT-1 in rat circumventricular organs, meninges, and dorsal root ganglia.
    J Comp Neurol. 2000 Jun 5;421(3):385-99 PMID: 10813794
  34. Total number and distribution of inhibitory and excitatory synapses on hippocampal CA1 pyramidal cells.
    Neuroscience. 2001;102(3):527-40 PMID: 11226691
  35. Glutamate uptake.
    Prog Neurobiol. 2001 Sep;65(1):1-105 PMID: 11369436
  36. A splice variant of glutamate transporter GLT1/EAAT2 expressed in neurons: cloning and localization in rat nervous system.
    Neuroscience. 2002;109(1):45-61 PMID: 11784699
  37. Distribution of two splice variants of the glutamate transporter GLT1 in the retinas of humans, monkeys, rabbits, rats, cats, and chickens.
    J Comp Neurol. 2002 Mar 25;445(1):1-12 PMID: 11891650
  38. Expression of a variant form of the glutamate transporter GLT1 in neuronal cultures and in neurons and astrocytes in the rat brain.
    J Neurosci. 2002 Mar 15;22(6):2142-52 PMID: 11896154
  39. Distribution of two splice variants of the glutamate transporter GLT-1 in rat brain and pituitary.
    Glia. 2002 May;38(3):246-55 PMID: 11968062
  40. Distribution of two splice variants of the glutamate transporter GLT-1 in the developing rat retina.
    J Comp Neurol. 2002 Jun 10;447(4):323-30 PMID: 11992519
  41. Protein kinase C activation decreases cell surface expression of the GLT-1 subtype of glutamate transporter. Requirement of a carboxyl-terminal domain and partial dependence on serine 486.
    J Biol Chem. 2002 Nov 29;277(48):45741-50 PMID: 12324450
  42. The 'glial' glutamate transporter, EAAT2 (Glt-1) accounts for high affinity glutamate uptake into adult rodent nerve endings.
    J Neurochem. 2003 Feb;84(3):522-32 PMID: 12558972
  43. Morphine withdrawal increases glutamate uptake and surface expression of glutamate transporter GLT1 at hippocampal synapses.
    J Neurosci. 2003 Jun 1;23(11):4775-84 PMID: 12805317
  44. High affinity glutamate transport in rat cortical neurons in culture.
    Mol Pharmacol. 1998 Jan;53(1):88-96 PMID: 9443935
  45. Extrasynaptic glutamate diffusion in the hippocampus: ultrastructural constraints, uptake, and receptor activation.
    J Neurosci. 1998 May 1;18(9):3158-70 PMID: 9547224
  46. Neuronal expression of the glutamate transporter GLT-1 in hippocampal microcultures.
    J Neurosci. 1998 Jun 15;18(12):4490-9 PMID: 9614226
  47. Glutamate transporter GLT-1 is transiently localized on growing axons of the mouse spinal cord before establishing astrocytic expression.
    J Neurosci. 1998 Aug 1;18(15):5706-13 PMID: 9671661
  48. Comparative analysis of glutamate transporter expression in rat brain using differential double in situ hybridization.
    Anat Embryol (Berl). 1998 Jul;198(1):13-30 PMID: 9683064
  49. Glial contribution to glutamate uptake at Schaffer collateral-commissural synapses in the hippocampus.
    J Neurosci. 1998 Oct 1;18(19):7709-16 PMID: 9742141
  50. Expression of the GLT-1 subtype of Na+-dependent glutamate transporter: pharmacological characterization and lack of regulation by protein kinase C.
    J Pharmacol Exp Ther. 1999 Jun;289(3):1600-10 PMID: 10336558
  51. GLT1, glial glutamate transporter, is transiently expressed in neurons and develops astrocyte specificity only after midgestation in the ovine fetal brain.
    J Neurobiol. 1999 Jun 15;39(4):515-26 PMID: 10380073
  52. Three-dimensional relationships between hippocampal synapses and astrocytes.
    J Neurosci. 1999 Aug 15;19(16):6897-906 PMID: 10436047
  53. High affinity uptake of glutamate in terminals of corticostriatal axons.
    Nature. 1977 Mar 24;266(5600):377-8 PMID: 859607
  54. Uptake of [3H]Glutamic acid in excitatory nerve endings: light and electronmicroscopic observations in the hippocampal formation of the rat.
    Neuroscience. 1979;4(9):1237-53 PMID: 492534
  55. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.
    J Histochem Cytochem. 1981 Apr;29(4):577-80 PMID: 6166661
  56. In vivo high-affinity uptake and axonal transport of D-[2,3-3H]aspartate in excitatory neurons.
    Brain Res. 1981 Dec 28;230(1-2):427-33 PMID: 6172187
  57. Chronic infusion of endogenous excitatory amino acids into rat striatum and hippocampus.
    Brain Res Bull. 1983 Jan;10(1):47-51 PMID: 6130829
  58. Glutamate: a neurotransmitter in mammalian brain.
    J Neurochem. 1984 Jan;42(1):1-11 PMID: 6139418
  59. Uptake of D-aspartate and L-glutamate in excitatory axon terminals in hippocampus: autoradiographic and biochemical comparison with gamma-aminobutyrate and other amino acids in normal rats and in rats with lesions.
    Neuroscience. 1984 Jan;11(1):79-100 PMID: 6143283
  60. Heterogeneity of sodium-dependent excitatory amino acid uptake mechanisms in rat brain.
    J Neurosci Res. 1986;16(3):491-503 PMID: 2877096
  61. A novel method for producing anti-peptide antibodies. Production of site-specific antibodies to the T cell antigen receptor beta-chain.
    J Biol Chem. 1988 Feb 5;263(4):1719-25 PMID: 3276675
  62. Synthetic peptide vaccine design: synthesis and properties of a high-density multiple antigenic peptide system.
    Proc Natl Acad Sci U S A. 1988 Aug;85(15):5409-13 PMID: 3399498
  63. Glutamine and 2-oxoglutarate as metabolic precursors of the transmitter pools of glutamate and GABA: correlation of regional uptake by rat brain synaptosomes.
    Neurochem Res. 1989 Apr;14(4):371-6 PMID: 2569675
  64. Optimization of differential immunogold-silver and peroxidase labeling with maintenance of ultrastructure in brain sections before plastic embedding.
    J Neurosci Methods. 1990 Aug;33(2-3):113-27 PMID: 1977960
  65. Non-vesicular release of glutamate from glial cells by reversed electrogenic glutamate uptake.
    Nature. 1990 Nov 29;348(6300):443-6 PMID: 2247147
  66. Glutamate uptake disguises neurotoxic potency of glutamate agonists in cerebral cortex in dissociated cell culture.
    J Neurosci. 1992 Jan;12(1):56-61 PMID: 1345946
  67. Complementary neuronal and glial expression of two high-affinity glutamate transporter GLT1/EAAT2 forms in rat cerebral cortex.
    Histochem Cell Biol. 2003 Jun;119(6):425-35 PMID: 12756574
  68. Functional changes of glial glutamate transporter GLT-1 during ischemia: an in vivo study in the hippocampal CA1 of normal mice and mutant mice lacking GLT-1.
    J Neurosci. 2003 Aug 6;23(18):7176-82 PMID: 12904478
  69. Localizing 3H-GABA in nerve terminals of rat cerebral cortex by electron microscopic autoradiography.
    Nature. 1971 Feb 26;229(5287):628-30 PMID: 4925465
  70. The autoradiographic localization of L-(3H) glutamate in synaptosomal preparations.
    Brain Res. 1976 Feb 20;103(2):350-5 PMID: 1252923
  71. Differential expression of two glial glutamate transporters in the rat brain: quantitative and immunocytochemical observations.
    J Neurosci. 1995 Mar;15(3 Pt 1):1835-53 PMID: 7891138
  72. Glutamate transporters in glial plasma membranes: highly differentiated localizations revealed by quantitative ultrastructural immunocytochemistry.
    Neuron. 1995 Sep;15(3):711-20 PMID: 7546749
  73. Immunocytochemical identification of cone bipolar cells in the rat retina.
    J Comp Neurol. 1995 Oct 23;361(3):461-78 PMID: 8550893
Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
Published
2004-02-04
Pages
1136-48
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC2849838
Subset
IM
Grants
NINDS NIH HHS · NS41883 · United States
NEI NIH HHS · P30 EY013079-069003 · United States
NINDS NIH HHS · R01 NS041091 · United States
NEI NIH HHS · P30 EY013079 · United States
NICHD NIH HHS · P30 HD018655 · United States
NEI NIH HHS · R01 EY013145 · United States
NINDS NIH HHS · NS40753 · United States
NINDS NIH HHS · R01 NS040753 · United States
NEI NIH HHS · R01 EY013145-04 · United States
NINDS NIH HHS · R01 NS041091-04 · United States
NINDS NIH HHS · NS41091 · United States
NEI NIH HHS · EY13145 · United States
NICHD NIH HHS · HD18655 · United States
NEI NIH HHS · EY13079 · United States
NINDS NIH HHS · R25 NS080686 · United States
NINDS NIH HHS · R01 NS041883 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com