title: Expression of the glial glutamate transporter EAAT2 in the human CNS: an immunohistochemical study.
abstract: Glutamate transporters play an essential role in terminating the excitatory glutamatergic signal at post-synaptic receptors and in protecting neurones from excitotoxic effects, as well as replenishing the neurotransmitter supply at glutamatergic synapses. The distribution and density of glutamate transporters may be important determinants of vulnerability to glutamate-mediated injury. There is emerging evidence that glutamate transporter dysfunction may be present in motor neurone disease (MND). In this study, a monoclonal antibody, suitable for immunohistochemistry (IHC) in human post-mortem tissue, was produced to the human astrocytic glutamate transporter EAAT2 (excitatory amino acid transporter 2). Western blotting of homogenates of human cortical tissue with the EAAT2 antibody produced a discrete band at 66 kDa. Detailed IHC analysis of the expression of the EAAT2 protein in the human CNS was undertaken. EAAT2 was exclusively localised to astrocytes, with preferential expression in the caudate nucleus, nucleus basalis of Meynert, spinal ventral horn, cerebral cortex and hippocampus, but with lower levels of expression throughout many other CNS regions. Motor neurone groups vulnerable to neurodegeneration in MND appeared distinctive in being surrounded by extensive, coarse, strongly immunoreactive perisomatic glial profiles. Motor neurone groups which tend to be spared in MND, such as those present in the oculomotor nucleus, showed a lower expression of EAAT2, with fewer perisomatic profiles. The EAAT2 antibody will provide a useful tool for increasing our understanding of the role of EAAT2 in excitatory neurotransmission in health and disease states.
authors.affiliation: Novocastra Laboratories, Newcastle upon Tyne, UK.
authors.forename: I D
authors.initials: ID
authors.lastname: Milton
authors.forename: S J
authors.initials: SJ
authors.lastname: Banner
authors.forename: P G
authors.initials: PG
authors.lastname: Ince
authors.forename: N H
authors.initials: NH
authors.lastname: Piggott
authors.forename: A E
authors.initials: AE
authors.lastname: Fray
authors.forename: N
authors.initials: N
authors.lastname: Thatcher
authors.forename: C H
authors.initials: CH
authors.lastname: Horne
authors.forename: P J
authors.initials: PJ
authors.lastname: Shaw
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000032:analysis*; D027322:Amino Acid Transport System X-AG; D000911:Antibodies, Monoclonal; D001479:Basal Ganglia / Q000737:chemistry; D001692:Biological Transport / Q000502:physiology; D015153:Blotting, Western; D002490:Central Nervous System / Q000166:cytology / Q000378:metabolism*; D002540:Cerebral Cortex / Q000737:chemistry; D006624:Hippocampus / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D008161:Lumbosacral Region; D018613:Microscopy, Confocal; D009044:Motor Cortex / Q000737:chemistry; D009457:Neuroglia / Q000378:metabolism*; D013116:Spinal Cord / Q000737:chemistry
chemical_list: D018528:ATP-Binding Cassette Transporters; D027322:Amino Acid Transport System X-AG; D000911:Antibodies, Monoclonal