title: Loss of glutamine synthetase in the human epileptogenic hippocampus: possible mechanism for raised extracellular glutamate in mesial temporal lobe epilepsy.
abstract: BACKGROUND
  High extracellular glutamate concentrations have been identified as a likely trigger of epileptic seizures in mesial temporal lobe epilepsy (MTLE), but the underlying mechanism remains unclear. We investigated whether a deficiency in glutamine synthetase, a key enzyme in catabolism of extracellular glutamate in the brain, could explain the perturbed glutamate homoeostasis in MTLE.
  
  
  METHODS
  The anteromedial temporal lobe is the focus of the seizures in MTLE, and surgical resection of this structure, including the hippocampus, leads to resolution of seizures in many cases. By means of immunohistochemistry, western blotting, and functional enzyme assays, we assessed the distribution, quantity, and activity of glutamine synthetase in the MTLE hippocampus.
  
  
  RESULTS
  In western blots, the expression of glutamine synthetase in the hippocampus was 40% lower in MTLE than in non-MTLE samples (median 44 [IQR 30-58] vs 69 [56-87]% of maximum concentration in standard curve; p=0.043; n=8 and n=6, respectively). The enzyme activity was lower by 38% in MTLE vs non-MTLE (mean 0.0060 [SD 0.0031] vs 0.0097 [0.0042] U/mg protein; p=0.045; n=6 and n=9, respectively). Loss of glutamine synthetase was particularly pronounced in areas of the MTLE hippocampus with astroglial proliferation, even though astrocytes normally have high content of the enzyme. Quantitative immunoblotting showed no significant change in the amount of EAAT2, the predominant glial glutamate transporter in the hippocampus.
  
  
  CONCLUSIONS
  A deficiency in glutamine synthetase in astrocytes is a possible molecular basis for extracellular glutamate accumulation and seizure generation in MTLE. Further studies are needed to define the cause, but the loss of glutamine synthetase may provide a new focus for therapeutic interventions in MTLE.
authors.affiliation: Department of Neurosurgery, Yale University School of Medicine, New Haven, CT, USA. tore.eid@yale.edu
authors.forename: T
authors.initials: T
authors.lastname: Eid
authors.forename: M J
authors.initials: MJ
authors.lastname: Thomas
authors.forename: D D
authors.initials: DD
authors.lastname: Spencer
authors.forename: E
authors.initials: E
authors.lastname: Rundén-Pran
authors.forename: J C K
authors.initials: JC
authors.lastname: Lai
authors.forename: G V
authors.initials: GV
authors.lastname: Malthankar
authors.forename: J H
authors.initials: JH
authors.lastname: Kim
authors.forename: N C
authors.initials: NC
authors.lastname: Danbolt
authors.forename: O P
authors.initials: OP
authors.lastname: Ottersen
authors.forename: N C
authors.initials: NC
authors.lastname: de Lanerolle
mesh_terms: D000293:Adolescent; D000328:Adult; D001253:Astrocytes / Q000201:enzymology / Q000378:metabolism; D015153:Blotting, Western; D002648:Child; D004833:Epilepsy, Temporal Lobe / Q000201:enzymology* / Q000378:metabolism; D027342:Excitatory Amino Acid Transporter 2 / Q000032:analysis / Q000378:metabolism; D005110:Extracellular Space / Q000737:chemistry / Q000378:metabolism; D005260:Female; D005974:Glutamate-Ammonia Ligase / Q000032:analysis* / Q000172:deficiency / Q000378:metabolism; D018698:Glutamic Acid / Q000032:analysis* / Q000378:metabolism; D006624:Hippocampus / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D013702:Temporal Lobe / Q000201:enzymology / Q000378:metabolism
chemical_list: D027342:Excitatory Amino Acid Transporter 2; D018698:Glutamic Acid; D005974:Glutamate-Ammonia Ligase