Glutamate clearance mechanisms in resected cortical dysplasia

Laboratory investigation

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Object

Changes in the expression of glutamate transporters (GLTs) may play a role in the expression of epileptogenicity. Previous studies have shown an increased number of neuronal GLTs in human dysplastic neurons. The expression of glial and neuronal GLTs and glutamine synthetase (GS) in balloon cells (BCs) and BC-containing cortical dysplasia has not been studied.

Methods

The authors analyzed neocortical samples that were resected in 5 patients who had cortical dysplasia–induced medically intractable focal epilepsy and who underwent extraoperative prolonged electrocorticographic (ECoG) recordings. The expressions of glial (GLT1/EAAT2) and neuronal (EAAT3, EAAC1) GLTs and GS proteins were immunohistochemically studied in all 5 resected samples. The authors also assessed in situ colocalization of GLTs and GS with neuronal and glial markers.

Results

Balloon cell–containing cortical dysplasia lesions did not exhibit ictal patterns on prolonged extraoperative ECoG recordings. There was a differential expression of glial and neuronal GLTs in BCs and dysplastic neurons: the majority of BCs highly expressed glial but not neuronal GLTs. Dysplastic neurons showed increased immunohistochemical staining with neuronal EAAT3 but not with EAAT2/GLT1. Moreover, only glial fibrillary acidic protein–positive BCs also expressed GS.

Conclusions

There is a differential GLT expression in dysplastic and balloon cells. The presence of glial GLTs and GS in balloon cell cortical dysplasia suggests a possible antiepileptic role for BCs and is consistent with the reported increased epileptogenicity in GLT1-deficient animals.

Abbreviations used in this paper: BC = balloon cell; ECoG = electrocorticographic; EEG = electroencephalography; FCD = focal cortical dysplasia; GFAP = glial fibrillary acidic protein; GLT = glutamate transporter; GS = glutamine synthetase; NeuN = neuronal nuclei; TBS = Tris-buffered saline.

Article Information

Address correspondence to: Jorge A. Gonzalez-Martinez, M.D., Ph.D., Epilepsy Center, Cleveland Clinic, 9500 Euclid Avenue, S51, Cleveland, Ohio 44195. email: gonzalj1@ccf.org.

Please include this information when citing this paper: published online November 12, 2010; DOI: 10.3171/2010.10.JNS10715.

© AANS, except where prohibited by US copyright law.

Headings

Figures

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    Imaging and electrophysiological and histopathological correlations. Upper: Axial FLAIR MR images showing an area of signal increase extending from the left precentral gyrus cortical mantle toward the periventricular region. On prolonged extraoperative direct ECoG recordings, the center of the lesion (A) did not show an in situ interictal or ictal epileptic patterns but exhibited pathological changes consistent with BC-containing cortical dysplasia (Type 2B [photomicrograph, lower left]). The surrounding cortex of the MR imaging visible lesion (B) showed evidence of in situ interictal and ictal epileptic patterns and exhibited pathological changes consistent with Type 2A cortical dysplasia (photomicrograph, lower right).

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    Immunocytochemical staining with EAAC1 antibody showing scattered and light staining of the BCs (A and B) and more dense and diffuse staining of the dysmorphic cells (C and D). Bar = 100 μm.

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    Double labeling of cells with MAP2 (green) (A) and neuronal GLT (EAAC1, red) (B) antibodies showing the colocalization of the 2 proteins (C). Bar = 100 μm.

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    Immunocytochemical staining with the 2 glial-specific GLT antibodies, EAAT2 (A and B) and GLT1 (C and D), showing diffuse and dense BC staining to both antibodies. Bar = 100 μm.

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    Double labeling of BCs with GFAP (green) (A) and glial glutamate transporter (EAT2, red) (B) antibodies showing the colocalization of the 2 proteins in BCs (C). Bar = 100 μm.

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    Immunocytochemical staining with GS antibodies showing astrocytic (A and B) and BC (C and D) staining. Bar = 100 μm.

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    Double labeling of BCs with GFAP (green, A) and GS (red, B) antibodies showing the colocalization of the 2 proteins in BCs (C). Bar = 100 μm.

References

  • 1

    Anderson CMSwanson RA: Astrocyte glutamate transport: review of properties, regulation, and physiological functions. Glia 32:1142000

    • Search Google Scholar
    • Export Citation
  • 2

    Bautista JFFoldvary-Schaefer NBingaman WELüders HO: Focal cortical dysplasia and intractable epilepsy in adults: clinical, EEG, imaging, and surgical features. Epilepsy Res 55:1311362003

    • Search Google Scholar
    • Export Citation
  • 3

    Binder DKSteinhäuser C: Functional changes in astroglial cells in epilepsy. Glia 54:3583682006

  • 4

    Bjørnsen LPEid THolmseth SDanbolt NCSpencer DDde Lanerolle NC: Changes in glial glutamate transporters in human epileptogenic hippocampus: inadequate explanation for high extracellular glutamate during seizures. Neurobiol Dis 25:3193302007

    • Search Google Scholar
    • Export Citation
  • 5

    Boonyapisit KNajm IKlem GYing ZBurrier CLaPresto E: Epileptogenicity of focal malformations due to abnormal cortical development: direct electrocorticographic-histopathologic correlations. Epilepsia 44:69762003

    • Search Google Scholar
    • Export Citation
  • 6

    Campbell SLHablitz JJ: Decreased glutamate transport enhances excitability in a rat model of cortical dysplasia. Neurobiol Dis 32:2542612008

    • Search Google Scholar
    • Export Citation
  • 7

    Carl GFBlackwell LKBarnett FCThompson LARissinger CJOlin KL: Manganese and epilepsy: brain glutamine synthetase and liver arginase activities in genetically epilepsy prone and chronically seizured rats. Epilepsia 34:4414461993

    • Search Google Scholar
    • Export Citation
  • 8

    Claudio OIFerchmin PVelísek LSperber EFMoshé SLOrtiz JG: Plasticity of excitatory amino acid transporters in experimental epilepsy. Epilepsia 41:Suppl 6S104S1102000

    • Search Google Scholar
    • Export Citation
  • 9

    Domercq MMatute C: Expression of glutamate transporters in the adult bovine corpus callosum. Brain Res Mol Brain Res 67:2963021999

    • Search Google Scholar
    • Export Citation
  • 10

    Dutuit MDidier-Bazès MVergnes MMutin MConjard AAkaoka H: Specific alteration in the expression of glial fibrillary acidic protein, glutamate dehydrogenase, and glutamine synthetase in rats with genetic absence epilepsy. Glia 32:15242000

    • Search Google Scholar
    • Export Citation
  • 11

    Furuta ARothstein JDMartin LJ: Glutamate transporter protein subtypes are expressed differentially during rat CNS development. J Neurosci 17:836383751997

    • Search Google Scholar
    • Export Citation
  • 12

    González-Martínez JABingaman WEToms SANajm IM: Neurogenesis in the postnatal human epileptic brain. J Neurosurg 107:6286352007

    • Search Google Scholar
    • Export Citation
  • 13

    Harrington EPMöddel GNajm IMBaraban SC: Altered glutamate receptor—transporter expression and spontaneous seizures in rats exposed to methylazoxymethanol in utero. Epilepsia 48:1581682007

    • Search Google Scholar
    • Export Citation
  • 14

    Ingram EMTessler SBowery NGEmson PC: Glial glutamate transporter mRNAs in the genetically absence epilepsy rat from Strasbourg. Brain Res Mol Brain Res 75:961042000

    • Search Google Scholar
    • Export Citation
  • 15

    Jeha LENajm IMBingaman WEKhandwala FWiddess-Walsh PMorris HH: Predictors of outcome after temporal lobectomy for the treatment of intractable epilepsy. Neurology 66:193819402006

    • Search Google Scholar
    • Export Citation
  • 16

    Kawashima TAdachi TTokunaga YFuruta ASuzuki SODoh-ura K: Immunohistochemical analysis in a case of idiopathic Lennox-Gastaut syndrome. Clin Neuropathol 18:2862921999

    • Search Google Scholar
    • Export Citation
  • 17

    Kondo KHashimoto HKitanaka JSawada MSuzumura AMarunouchi T: Expression of glutamate transporters in cultured glial cells. Neurosci Lett 188:1401421995

    • Search Google Scholar
    • Export Citation
  • 18

    Laming PRCosby SLO'Neill JK: Seizures in the Mongolian gerbil are related to a deficiency in cerebral glutamine synthetase. Comp Biochem Physiol C 94:3994041989

    • Search Google Scholar
    • Export Citation
  • 19

    Lamparello PBaybis MPollard JHol EMEisenstat DDAronica E: Developmental lineage of cell types in cortical dysplasia with balloon cells. Brain 130:226722762007

    • Search Google Scholar
    • Export Citation
  • 20

    Lüders HSchuele SU: Epilepsy surgery in patients with malformations of cortical development. Curr Opin Neurol 19:1691742006

  • 21

    Mathern GWMendoza DLozada APretorius JKDehnes YDanbolt NC: Hippocampal GABA and glutamate transporter immunoreactivity in patients with temporal lobe epilepsy. Neurology 52:4534721999

    • Search Google Scholar
    • Export Citation
  • 22

    Milh MBecq HVilleneuve NBen-Ari YAniksztejn L: Inhibition of glutamate transporters results in a “suppression-burst” pattern and partial seizures in the newborn rat. Epilepsia 48:1691742007

    • Search Google Scholar
    • Export Citation
  • 23

    Mischel PSNguyen LPVinters HV: Cerebral cortical dysplasia associated with pediatric epilepsy. Review of neuropathologic features and proposal for a grading system. J Neuropathol Exp Neurol 54:1371531995

    • Search Google Scholar
    • Export Citation
  • 24

    Najm IMBingaman WELüders HO: The use of subdural grids in the management of focal malformations due to abnormal cortical development. Neurosurg Clin N Am 13:8792viiiix2002

    • Search Google Scholar
    • Export Citation
  • 25

    Najm IMTilelli CQOghlakian R: Pathophysiological mechanisms of focal cortical dysplasia: a critical review of human tissue studies and animal models. Epilepsia 48:Suppl 221322007

    • Search Google Scholar
    • Export Citation
  • 26

    Najm IMYing ZBabb TMohamed AHadam JLaPresto E: Epileptogenicity correlated with increased N-methyl-D-aspartate receptor subunit NR2A/B in human focal cortical dysplasia. Epilepsia 41:9719762000

    • Search Google Scholar
    • Export Citation
  • 27

    Oh HSLee MCKim HSLee JSLee JHKim MK: Pathophysiologic characteristics of balloon cells in cortical dysplasia. Childs Nerv Syst 24:1751832008

    • Search Google Scholar
    • Export Citation
  • 28

    O'Kane RLMartínez-López IDeJoseph MRViña JRHawkins RA: Na(+)-dependent glutamate transporters (EAAT1, EAAT2, and EAAT3) of the blood-brain barrier. A mechanism for glutamate removal. J Biol Chem 274:31891318951999

    • Search Google Scholar
    • Export Citation
  • 29

    Palmini ANajm IAvanzini GBabb TGuerrini RFoldvary-Schaefer N: Terminology and classification of the cortical dysplasias. Neurology 62:6 Suppl 3S2S82004

    • Search Google Scholar
    • Export Citation
  • 30

    Rothstein JDDykes-Hoberg MPardo CABristol LAJin LKuncl RW: Knockout of glutamate transporters reveals a major role for astroglial transport in excitotoxicity and clearance of glutamate. Neuron 16:6756861996

    • Search Google Scholar
    • Export Citation
  • 31

    Rothstein JDMartin LLevey AIDykes-Hoberg MJin LWu D: Localization of neuronal and glial glutamate transporters. Neuron 13:7137251994

    • Search Google Scholar
    • Export Citation
  • 32

    Scimemi ASchorge SKullmann DMWalker MC: Epileptogenesis is associated with enhanced glutamatergic transmission in the perforant path. J Neurophysiol 95:121312202006

    • Search Google Scholar
    • Export Citation
  • 33

    Stafstrom CE: The role of glutamate transporters in developmental epilepsy: a concept in flux. Epilepsy Curr 4:2432442004

  • 34

    Tanaka KWatase KManabe TYamada KWatanabe MTakahashi K: Epilepsy and exacerbation of brain injury in mice lacking the glutamate transporter GLT-1. Science 276:169917021997

    • Search Google Scholar
    • Export Citation
  • 35

    Tassi LColombo NGarbelli RFrancione SLo Russo GMai R: Focal cortical dysplasia: neuropathological subtypes, EEG, neuroimaging and surgical outcome. Brain 125:171917322002

    • Search Google Scholar
    • Export Citation
  • 36

    Tassi LPasquier BMinotti LGarbelli RKahane PBenabid AL: Cortical dysplasia: electroclinical, imaging, and neuropathologic study of 13 patients. Epilepsia 42:111211232001

    • Search Google Scholar
    • Export Citation
  • 37

    Tiffany-Castiglioni ECPeterson SLCastiglioni AJ: Alterations in glutamine synthetase activity by FeCl2-induced focal and kindled amygdaloid seizures. J Neurosci Res 25:2232281990

    • Search Google Scholar
    • Export Citation
  • 38

    Ueda YWillmore LJ: Sequential changes in glutamate transporter protein levels during Fe(3+)-induced epileptogenesis. Epilepsy Res 39:2012092000

    • Search Google Scholar
    • Export Citation
  • 39

    van der Hel WSVerlinde SAMeijer DHde Wit MRensen MGvan Gassen KL: Hippocampal distribution of vesicular glutamate transporter 1 in patients with temporal lobe epilepsy. Epilepsia 50:171717282009

    • Search Google Scholar
    • Export Citation
  • 40

    Widdess-Walsh PJeha LNair DKotagal PBingaman WNajm I: Subdural electrode analysis in focal cortical dysplasia: predictors of surgical outcome. Neurology 69:6606672007

    • Search Google Scholar
    • Export Citation
  • 41

    Ying ZGonzalez-Martinez JTilelli CBingaman WNajm I: Expression of neural stem cell surface marker CD133 in balloon cells of human focal cortical dysplasia. Epilepsia 46:171617232005

    • Search Google Scholar
    • Export Citation

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