Journal of Biomedical Science | |
Rapid compensatory changes in the expression of EAAT-3 and GAT-1 transporters during seizures in cells of the CA1 and dentate gyrus | |
Silvia J López-Pérez1  Alberto Morales-Villagrán1  Flavio Sandoval-García1  Laura Medina-Ceja1  | |
[1] Laboratorio de Neurofisiología y Neuroquímica, Departamento de Biología Celular y Molecular, Centro Universitario de Ciencias Biológicas y Agropecuarias, Universidad de Guadalajara, Km. 15.5 Carretera Guadalajara-Nogales Predio “Las Agujas”; Nextipac, Zapopan, Jalisco, CP 45110, Mexico | |
关键词: Seizures; Immunofluorescence; Hippocampus; GAT-1; EAAT-3; 4-Aminopyridine; | |
Others : 824607 DOI : 10.1186/1423-0127-19-78 |
|
received in 2012-04-04, accepted in 2012-08-21, 发布年份 2012 | |
【 摘 要 】
Background
Epilepsy is a neurological disorder produced by an imbalance between excitatory and inhibitory neurotransmission, in which transporters of both glutamate and GABA have been implicated. Hence, at different times after local administration of the convulsive drug 4-aminopyridine (4-AP) we analyzed the expression of EAAT-3 and GAT-1 transporter proteins in cells of the CA1 and dentate gyrus.
Methods
Dual immunofluorescence was used to detect the co-localization of transporters and a neuronal marker. In parallel, EEG recordings were performed and convulsive behavior was rated using a modified Racine Scale.
Results
By 60 min after 4-AP injection, EAAT-3/NeuN co-labelling had increased in dentate granule cells and decreased in CA1 pyramidal cells. In the latter, this decrease persisted for up to 180 min after 4-AP administration. In both the DG and CA1, the number of GAT-1 labeled cells increased 60 min after 4-AP administration, although by 180 min GAT-1 labeled cells decreased in the DG alone. The increase in EAAT-3/NeuN colabelling in DG was correlated with maximum epileptiform activity and convulsive behavior.
Conclusions
These findings suggest that a compensatory mechanism exists to protect against acute seizures induced by 4-AP, whereby EAAT-3/NeuN cells is rapidly up regulated in order to enhance the removal of glutamate from the extrasynaptic space, and attenuating seizure activity.
【 授权许可】
2012 Medina-Ceja et al.; licensee BioMed Central Ltd.
【 预 览 】
Files | Size | Format | View |
---|---|---|---|
20140713040223425.pdf | 1958KB | download | |
Figure 4. | 49KB | Image | download |
Figure 3. | 138KB | Image | download |
Figure 2. | 68KB | Image | download |
Figure 1. | 84KB | Image | download |
【 图 表 】
Figure 1.
Figure 2.
Figure 3.
Figure 4.
【 参考文献 】
- [1]Angus-Leppan H, Parsons LM: Epilepsy: epidemiology, classification and natural history. Medicine 2008, 11:571-578.
- [2]Medina-Ceja L, Morales-Villagrán A, Tapia R: Action of 4-aminopyridine on extracellular amino acids in hippocampus and entorhinal cortex: a dual microdialysis and electroencephalographic study in awake rats. Brain Res Bull 2000, 53:255-262.
- [3]Tapia R, Medina-Ceja L, Peña F: On the relationship between extracellular glutamate, hyperexcitation and neurodegeneration, in vivo. Neurochem Int 1999, 34:23-31.
- [4]Morales-Villagrán A, Medina-Ceja L, López-Pérez SJ: Simultaneous glutamate and EEG activity measurements during seizures in rat hippocampal región with the use of an electrochemical biosensor. J Neurosci Methods 2008, 168:48-53.
- [5]Lauriat TL, McInnes LA: EAAT-2 regulation and splicing: relevance to psychiatric and neurological disorders. Mol Psychiatry 2007, 12:1065-1078.
- [6]Medina-Ceja L, Guerrero-Cazares H, Canales-Aguirre A, Morales-Villagran A, Feria-Velasco A: Structural and functional characteristics of glutamate transporters: how they are related to epilepsy and oxidative stress. Rev Neurol 2007, 45:341-352.
- [7]Melone M, Bellesi M, Ducati A, Lacoangeli M, Conti F: Cellular and synaptic localization of EAAT2a in human cerebral cortex. Front Neuroanat 2011, 4:151.
- [8]Furuta A, Martin LJ, Lin CL, Dykes-Hoberg M, Rothstein JD: Cellular and synaptic localization of the neuronal glutamate transporters excitatory amino acid transporter 3 and 4. Neuroscience 1997, 81:1031-1042.
- [9]Maragakis NJ, Rothstein JD: Glutamate transporters in neurologic disease. Arch Neurol 2001, 58:365-370.
- [10]Rothstein JD, Martin L, Levey AI, Dykes-Hoberg M, Jin L, Wu D, Nash N, Kuncl RW: Localization of neuronal and glial glutamate transporters. Neuron 1994, 13:713-725.
- [11]Lewerenz J, Klein M, Methner A: Cooperative action of glutamate transporters and cystine/glutamate antiporter system Xc- protects from oxidative glutamate toxicity. J Neurochem 2006, 98:916-925.
- [12]Crino PB, Jin H, Shumate MD, Robinson MB, Coulter DA, Brooks-Kayal AR: Increased Expression of the Neuronal Glutamate Transporter (EAAT3/EAAC1) in Hippocampal and Neocortical Epilepsy. Epilepsia 2002, 43:211-218.
- [13]Mathern GW, Mendoza D, Lozada A, Peerorius JK, Dehnes Y, Danbolt NC, Nelson N, Leite JP, Chimelli L, Born DE, Sakamoto AC, Assirati JA, Fried I, Peacock WJ, Ojemann GA, Adelson PD: Hippocampal GABA and glutamate transporter immunoreactivity in patients with temporal lobe epilepsy. Neurology 1999, 52:453-472.
- [14]Proper EA, Hoogland G, Kappen SM, Jansen GH, Rensen MG, Schrama LH, van Veelen CW, van Rijen PC, van Nieuwenhuizen O, Gispen WH, de Graan PN: Distribution of glutamate transporters in the hippocampus of patients with pharmaco-resistant temporal lobe epilepsy. Brain 2002, 125:32-43.
- [15]Zhang G, Raol YSH, Hsu F-C, Brooks-Kayal AR: Long-term alterations in glutamate receptor and transporter expression following early-life seizures are associated with increased seizure susceptibility. J Neurochem 2004, 88:91-101.
- [16]Ueda Y, Willmore LJ: Hippocampal gamma-aminobutyric acid transporter alterations following focal epileptogenesis induced in rat amygdala. Brain Res Bull 2000, 52:357-361.
- [17]Sarup A, Larsson OM, Schousboe A: GABA transporters and GABA-transaminase as drug targets. Curr Drug Targets CNS Neurol Disord 2003, 2:269-277.
- [18]Yan XX, Carriaga WA, Ribak CE: Immunoreactivity for GABA plasma membrane transporter, GAT-1, in the developing rat cerebral cortex: transient presence in the somata of neocortical and hippocampal neurons. Develop Brain Res 1997, 99:1-19.
- [19]Jiang KW, Gao F, Shui QX, Yu ZS, Xia ZZ: Effect of diazoxide on regulation of vesicular and plasma membrane GABA transporter genes and proteins in hippocampus of rats subjected to picrotoxin-induced kindling. Neurosci Res 2004, 50:319-329.
- [20]Lee TS, Bjornsen LP, Paz C, Kim JH, Spencer SS, Spencer DD, Eid T, de Lanerolle NC: GAT-1 and GAT-3 expression are differently localized in the human epileptigenic hippocampus. Acta Neuropathol 2006, 111:351-363.
- [21]Sperk G, Schwarzer C, Heilman J, Furtinger S, Reimer RJ, Edwards RH, Nelson N: Expression of plasma membrane GABA transporters but not of the vesicular GABA transporter in dentate granule cells after kainic acid seizures. Hippocampus 2003, 13:806-15.
- [22]Peña F, Tapia R: Relationships among seizures, extracellular amino acids changes, and neurodegeneration induced by 4-aminopyridine in rat hippocampus: A microdialysis and electroencephalographic study. J Neurochem 1999, 72:2006-2014.
- [23]Medina-Ceja L, Cordero-Romero A, Morales-Villagrán A: Antiepileptic effect of carbenoxolone on seizures induced by 4-aminopyridine: A study in the rat hippocampus and entorhinal cortex. Brain Res 2008, 1187:74-81.
- [24]Medina-Ceja L, Ventura-Mejía C: Differential effects of trimethylamine and quinine on seizures induced by 4-aminopyridine administration in the entorhinal cortex of vigilant rats. Seizure 2010, 19:507-513.
- [25]Medina-Ceja L, Sandoval-García F, Pardo-Peña K: Effect of early glutamate exposure on EAAT-3 and GAT-1 protein expression in cells of the dentate gyrus and CA1 region of the Adult Rat Hippocampus. Arch Med Res 2011, 42:433-438.
- [26]West MJ: New Stereological methods for counting neurons. Neurobiol Aging 1993, 14:275-285.
- [27]Holmseth S, Dehnes Y, Huang YH, Follin-Arbelet VV, Grutle NJ, Mylonakou MN, Plachez C, Zhou Y, Furness DN, Bergles DE, Lehre KP, Danbolt NC: The density of EAAC1 (EAAT3) glutamate transporters expressed by neurons in the mammalian CNS. J Neurosci 2012, 32:6000-6013.
- [28]Simantov R, Crispino M, Hoe W, Broutman G, Tocco G, Rothstein JD, Baudry M: Changes in expression of neuronal and glial glutamate transporters in rat hippocampus following kainate-induced seizure activity. Brain Res Mol Brain Res 1999, 65:112-123.
- [29]Gorter JA, Van Vliet EA, Proper EA, De Gran PN, Ghijsen WE, Lopes Da Silva FH, Aronica E: Glutamate transporters alterations in the reorganizing dentate gyrus are associated with progressive seizure activity in chronic epileptic rats. J Comp Neurol 2002, 442:365-377.
- [30]Hanaya R, Sasa M, Sugata S, Tokudome M, Serikawa T, Kurisu K, Arita K: Hippocampal cell loss and propagation of abnormal discharges accompanied with the expression of tonic convulsion in the spontaneously epileptic rat. Brain Res 2010, 1328:171-180.
- [31]Qian J, Xu K, Yoo J, Chen TT, Andrews G, Noebels JL: Knockout of Zn transporters Zip-1 and Zip-3 attenuates seizure-induced CA1 neurodegeneration. J Neuroci 2011, 31:97-104.
- [32]Peña F, Tapia R: Seizures and neurodegeneration induced by 4-aminopyridine in rat hippocampus in vivo: Role of glutamate, and GABA mediated neurotransmition and of ion channels. Neuroscience 2000, 101:547-561.
- [33]Sakurai SY, Cha JH, Penney JB, Young AB: Regional distribution and properties of [3 H]MK-801 binding sites determined by quantitative autoradiography in rat brain. Neuroscience 1991, 40:533-543.
- [34]Morales-Villagrán A, Ureña-Guerrero M, Tapia R: Protection by NMDA receptor antagonists against seizures induced by intracerebral administration of 4-aminopyridine. Eur J Pharmacol 1996, 305:87-93.
- [35]Bagetta G, Iannone M, Palma E, Nisticó G, Dolly JO: N-methyl-D-aspartate and non-N-methyl-D-aspartate receptors mediate seizures and CA1 hippocampal damage induced by dendrotoxin-K in rats. Neuroscience 1996, 71:613-624.
- [36]Fournier KM, González MI, Robinson MB: Rapid trafficking of the neuronal glutamate transporter, EAAC1: evidence for distinct trafficking pathways differentially regulated by protein kinase C and platelet-derived growth factor. J Biol Chem 2004, 279:34505-34513.
- [37]Sheldon AI, González MI, Robinson MB: A carboxyl-terminal determinant of the neuronal glutamate transporter, EAAC1, is required for platelet-derived growth factor-dependent trafficking. J Biol Chem 2006, 281:4876-4886.
- [38]D’Amico A, Soragna A, Di Cairano E, Panzeri N, Anzai N, Vellea Sacchi F, Perego C: The surface density of the glutamate transporter EAAC1 is controlled by interactions with PDZK1 and AP2 adaptor complexes. Traffic 2010, 11:1455-1470.
- [39]Ross JR, Porter BE, Buckley PT, Eberwine JH, Robinson MB: mRNA for the EAAC1 subtype of glutamate transporter is present in neuronal dendrites in vitro and dramatically increases in vivo after seizure. Neurochem Int 2011, 58:366-375.
- [40]Morales-Villagrán A, Sandoval-Salazar C, Medina-Ceja L: An analytical flow injection system to measure glutamate in microdialysis samples based on an enzymatic reaction and electrochemical detection. Neurochem Res 2008, 33:1592-1598.
- [41]Zuo Z, Fang H: Glutamate transporter type 3 attenuates the activation of N-methyl-D-aspartate receptors co-expressed in Xenopus oocytes. J Exp Biol 2005, 208:2063-2070.
- [42]Schwarzer C, Sperk G: Hippocampal granule cells express glutamic acid decarboxylase-67 after limbic seizures in the rat. Neuroscience 1995, 69:705-709.
- [43]Sloviter RS, Dichter MA, Rachinsky TL, Dean E, Goodman JH, Sollas AL, Martin DL: Basal expression and induction of glutamate decarboxylase and GABA in excitatory granule cells of the rat and monkey hippocampal dentate gyrus. J Comp Neurol 1996, 373:593-618.
- [44]Mathews GC, Diamond JS: Neural glutamate uptake contributes to GABA synthesis and inhibitory synaptic strength. J Neurosci 2003, 23:2040-2048.
- [45]Sepkuty JP, Cohen AS, Eccles C, Rafiq A, Behar K, Ganel R, Coulter DA, Rothsein JD: A neuronal glutamate transporter contributes to neurotransmitter GABA synthesis and epilepsy. J Neurosci 2002, 22:6372-6379.
- [46]Gutierrez R: Seizures induce simultaneous GABAergic and glutamatergic transmission in the dentate gyrus-CA3 system. J Neurophysiol 2000, 84:3088-3090.
- [47]Doi T, Ueda Y, Tokumaru J, Mitsuyama Y, Willmore LJ: Sequential changes in glutamate transporter mRNA levels during Fe3+ −induced epileptogenesis. Mol Brain Res 2000, 75:105-112.
- [48]Rothstein JD, Van Kammen M, Levey AI, Martin LJ, Kuncl RW: Selective loss of glial glutamate transporter GLT-1 in amyotrophic lateral sclerosis. Ann Neurol 1995, 38:73-84.
- [49]Siniscalchi A, Calabresi P, Mercuri NB, Bernardi G: Epileptiform discharge induced by 4-aminopyridine in magnesium-free medium in neocortical neurons: physiological and pharmacological characterization. Neuroscience 1997, 81:189-197.
- [50]Young AB, Sakurai SY, Albin RL, Makowiec R, Penney JB: Excitatory amino acid receptor distribution: quantitative autoradiographic studies. In Excitatory Amino Acids and Synaptic Transmission. Edited by Thompson AW. Academic, USA; 1991:19-31.
- [51]Scimemi A, Tian H, Diamond JS: Neuronal transporters regulate glutamate clearance, NMDA receptor activation, and synaptic plasticity in the hippocampus. J Neurosci 2009, 29:14581-14595.
- [52]Ribak CE, Tong WM, Brecha NC: GABA plasma membrane transporters, GAT-1 and GAT-3, display different distributions in the rat hippocampus. J Comp Neurol 1996, 367:595-606.
- [53]Hirao T, Morimoto K, Yamamoto Y, Watanabe T, Sato H, Sato K, Sato S, Yamada M, Tanaka K, Suwaki H: Time-dependent and regional expression of GABA transporter mRNAs following amygdala-kindled seizures in rats. Brain Res Mol Brain Res 1998, 54:49-55.
- [54]Wu Y, Wang W, Díez-Sampedro A, Richerson GB: Nonvesicular inhibitory neurotransmission via reversal of the GABA transporter GAT-1. Neuron 2007, 56:851-865.
- [55]Richerson GB, Yuanming W: Dynamic equilibrium of neurotransmitter transporters: not just for reuptake anymore. J Neurophisiol 2003, 90:1363-1374.
- [56]Barakat L, Bordey A: GAT-1 and reversible GABA transport in Bergmann glia in slices. J Neurophysiol 2002, 88:1407-1417.
- [57]Salazar P, Tapia R: Allopregnanolone potentiates the glutamate-mediated seizures induced by 4-aminopyridine in rat hippocampus in vivo. Neurochem Res 2012, 37:596-603.