Amino Acids

, Volume 40, Issue 1, pp 221–238 | Cite as

Characterization, using comparative proteomics, of differentially expressed proteins in the hippocampus of the mesial temporal lobe of epileptic rats following treatment with valproate

  • Liwen Wu
  • Jing Peng
  • Chaoping Wei
  • Gu Liu
  • Guoli Wang
  • Kongzhao Li
  • Fei Yin
Original Article

Abstract

The objective of the study was to explore the pathogenesis of mesial temporal lobe epilepsy (MTLE) and the mechanism of valproate administration in the early stage of MTLE development. We performed a global comparative analysis and function classification of differentially expressed proteins using proteomics. MTLE models of developmental rats were induced by lithium-pilocarpine. Proteins in the hippocampus were separated by 2-DE technology. PDQuest software was used to analyze 2-DE images, and MALDI-TOF-MS was used to identify the differentially expressed proteins. Western blot was used to determine the differential expression levels of synapse-related proteins synapsin-1, dynamin-1 and neurogranin in both MTLE rat and human hippocampus. A total of 48 differentially expressed proteins were identified between spontaneous and non-spontaneous MTLE rats, while 41 proteins between MTLE rats and post valproate-treatment rats were identified. All of the proteins can be categorized into several groups by biological functions: synaptic and neurotransmitter release, cytoskeletal structure and dynamics, cell junctions, energy metabolism and mitochondrial function, molecular chaperones, signal regulation and others. Western blot results were similar to the changes noted in 2-DE. The differentially expressed proteins, especially the proteins related to synaptic and neurotransmitter release function, such as synapsin-1, dynamin-1 and neurogranin, are probably involved in the mechanism of MTLE and the pharmacological effect of valproate. These findings may provide important clues to elucidate the mechanism of chronic MTLE and to identify an optimum medication intervention time and new biomarkers for the development of pharmacological therapies targeted at epilepsy.

Keywords

Mesial temporal lobe epilepsy Hippocampus Valproate Proteomics Mechanism 

Abbreviations

AEDs

Antiepileptic drugs

Cx50

Gap junction channel protein connexin 50

2-DE

Two-dimensional polyacrylamide gel electrophoresis

F-actin

Actin filaments

HSPs

Heat shock proteins

IML

Inner molecular layer

MALDI-TOF-MS

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

MAPKK1

Dual specificity mitogen-activated protein kinase kinase 1

MTLE

Mesial temporal lobe epilepsy

SE

Status epilepticus

SIR2-like

NAD-dependent deacetylase sirtuin-2

TCP-1

T-complex protein 1

VPA

Valproate

Notes

Acknowledgments

We thank Dr. Zhiquan Yang (Department of Neurosurgery, Xiangya Hospital, China) for providing the experimental hippocampus of MTLE patients and Dr. Zhaojun Duan (Department of Clinical Laboratory, Xiangya Hospital, China) for critically reviewing the manuscript. This work was supported by the National Natural Science Foundation of China, 30901631. The authors declare that there is no conflict of interest in either financial support or relationships.

References

  1. Babb TL, Kupfer WR, Pretorius JK, Crandall PH, Levesque MF (1991) Synaptic reorganization by mossy fibers in human epileptic fascia dentata. Neuroscience 42(2):351–363CrossRefPubMedGoogle Scholar
  2. Bakolitsa C, Cohen DM, Bankston LA, Bobkov AA, Cadwell GW, Jennings L, Critchley DR, Craig SW, Liddington RC (2004) Structural basis for vinculin activation at sites of cell adhesion. Nature 430(6999):513–515CrossRefGoogle Scholar
  3. Cavazos JE, Cross DJ (2006) The role of synaptic reorganization in mesial temporal lobe epilepsy. Epilepsy Behav 8(3):483–493CrossRefPubMedGoogle Scholar
  4. Danzer SC, He X, Loepke AW, McNamara JO (2010) Structural plasticity of dentate granule cell mossy fibers during the development of limbic epilepsy. Hippocampus 20(1):113–124PubMedGoogle Scholar
  5. Demand J, Luders J, Hoehfeld J (1998) The carboxy-terminal domain of Hsc70 provides binding sites for a distinct set of chaperone cofactors. Mol Cell Biol 18(4):2023–2028PubMedGoogle Scholar
  6. Dryden SC, Nahhas FA, Nowak JE, Goustin AS, Tainsky MA (2003) Role for human SIRT2 NAD-dependent deacetylase activity in control of mitotic exit in the cell cycle. Mol Cell Biol 23(9):3173–3185CrossRefPubMedGoogle Scholar
  7. Engel J Jr (1996) Introduction to temporal lobe epilepsy. J Epilepsy Res 26(1):141–150CrossRefGoogle Scholar
  8. Evergren E, Tomilin N, Vasylieva E, Sergeeva V, Bloom O, Gad H, Capani F, Shupliakov O (2004) A pre-embedding immunogold approach for detection of synaptic endocytic proteins in situ. J Neurosci Methods 135(1–2):169–174CrossRefPubMedGoogle Scholar
  9. Fioravante D, Liu RY, Netek AK, Cleary LJ, Byrne JH (2007) Synapsin regulates basal synaptic strength, synaptic depression, and serotonin-induced facilitation of sensorimotor synapses in Aplysia. J Neurophysiol 98(6):3568–3580CrossRefPubMedGoogle Scholar
  10. Franck JE, Pokorny J, Kunkel DD, Schwartzkroin PA (1995) Physiologic and morphologic characteristics of granule cell circuitry in human epileptic hippocampus. Epilepsia 36(6):543–558CrossRefPubMedGoogle Scholar
  11. Fukata Y, Adesnik H, Iwanaga T, Bredt DS, Nicoll RA, Fukata M (2006) Epilepsy-related ligand/receptor complex LGI1 and ADAM22 regulate synaptic transmission. Science 313(5794):1792–1795CrossRefPubMedGoogle Scholar
  12. Gao Y, Tatavarty V, Korza G, Levin MK, Carson JH (2008) Multiplexed dendritic targeting of alpha calcium calmodulin-dependent protein kinase II, neurogranin, and activity-regulated cytoskeleton-associated protein RNAs by the A2 pathway. Mol Biol Cell 19(5):2311–2327CrossRefPubMedGoogle Scholar
  13. Gluck MR, Jayatilleke E, Shaw S, Rowan AJ, Haroutunian V (2000) CNS oxidative stress associated with the kainic acid rodent model of experimental epilepsy. Epilepsy Res 39(1):63–71CrossRefPubMedGoogle Scholar
  14. Graves TD (2006) Ion channels and epilepsy. QJM 99(4):201–217CrossRefPubMedGoogle Scholar
  15. Greene ND, Bamidele A, Choy M, de Castro SC, Wait R, Leung KY, Begum S, Gadian DG, Scott RC, Lythgoe MF (2007) Proteome changes associated with hippocampal MRI abnormalities in the lithium pilocarpine-induced model of convulsive status epilepticus. Proteomics 7(8):1336–1344CrossRefPubMedGoogle Scholar
  16. Hamani C, Paulo I, Mello LE (2005) Neo-Timm staining in the thalamus of chronically epileptic rats. Braz J Med Biol Res 38(11):1677–1682CrossRefPubMedGoogle Scholar
  17. Hilfiker S, Benfenati F, Doussau F, Nairn AC, Czernik AJ, Augustine GJ, Greengard P (2005) Structural domains involved in the regulation of transmitter release by synapsins. J Neurosci 25(10):2658–2669CrossRefPubMedGoogle Scholar
  18. Hoeffer CA, Sanyal S, Ramaswami M (2003) Acute induction of conserved synaptic signaling pathways in Drosophila melanogaster. J Neurosci 23(15):6362–6372PubMedGoogle Scholar
  19. Horne MM, Guadagno TM (2003) A requirement for MAP kinase in the assembly and maintenance of the mitotic spindle. J Cell Biol 161(6):1021–1028CrossRefPubMedGoogle Scholar
  20. Huang KP, Huang FL, Ger JT, Li J, Reymann KG, Balschun D (2004) Neurogranin/RC3 enhances long-term potentiation and learning by promoting calcium-mediated signaling. J Neurosci 24(47):10660–10669CrossRefPubMedGoogle Scholar
  21. Jabs R, Seifert G, Steinh user C (2008) Astrocytic function and its alteration in the epileptic brain. Epilepsia 49(2):3–12CrossRefPubMedGoogle Scholar
  22. Johannessen CU, Petersen D, Fonnum F, Hassel B (2001) The acute effect of valproate on cerebral energy metabolism in mice. Epilepsy Res 47(3):247–256CrossRefPubMedGoogle Scholar
  23. Kapur J (2008) Is epilepsy a disease of synaptic transmission? Epilepsy Curr 8(5):139–141CrossRefPubMedGoogle Scholar
  24. Kubota Y, Putkey JA, Shouval HZ, Waxham MN (2008) IQ-motif proteins influence intracellular free Ca2+ in hippocampal neurons through their interactions with calmodulin. J Neurophysiol 99(1):264–276CrossRefPubMedGoogle Scholar
  25. Kwan P, Brodie MJ (2000) Epilepsy after the first drug fails: substitution or add-on? Seizure 9(7):464–468CrossRefPubMedGoogle Scholar
  26. Landmark CJ (2007) Targets for antiepileptic drugs in the synapse. Med Sci Monit 13(1):RA1–RA7PubMedGoogle Scholar
  27. Lee S, Carson K, Rice-Ficht A, Good T (2005) Hsp20, a novel alpha-crystallin, prevents Abeta fibril formation and toxicity. Protein Sci 14(3):593–601CrossRefPubMedGoogle Scholar
  28. Leite JP, Cavalheiro EA (1995) Effects of conventional antiepileptic drugs in a model of spontaneous recurrent seizures in rats. Epilepsy Res 20(2):93–104CrossRefPubMedGoogle Scholar
  29. Little E, Tocco G, Baudry M, Lee AS, Schreiber SS (1996) Induction of glucose-regulated protein (glucose-regulated protein 78/BiP and glucose-regulated protein 94) and heat shock protein 70 transcripts in the immature rat brain following status epilepticus. Neuroscience 75(1):209–219CrossRefPubMedGoogle Scholar
  30. Liu XY, Yang JL, Chen LJ, Zhang Y, Yang ML, Wu YY, Li FQ, Tang MH, Liang SF, Wei YQ (2008) Comparative proteomics and correlated signaling network of rat hippocampus in the pilocarpine model of temporal lobe epilepsy. Proteomics 8(3):582–603CrossRefPubMedGoogle Scholar
  31. Lopantsev V, Both M, Draguhn A (2009) Rapid plasticity at inhibitory and excitatory synapses in the hippocampus induced by ictal epileptiform discharges. Eur J Neurosci 29(6):1153–1164CrossRefPubMedGoogle Scholar
  32. Mathern GW, Babb TL, Micevych PE, Blanco CE, Pretorius JK (1997) Granule cell mRNA levels for BDNF, NGF, and NT-3 correlate with neuron losses or supragranular mossy fiber sprouting in the chronically damaged and epileptic human hippocampus. Mol Chem Neuropathol 30(1–2):53–76CrossRefPubMedGoogle Scholar
  33. McPherson PS, Czernik AJ, Chilcote TJ, Onofri F, Benfenati F, Greengard P, Schlessinger J, De Camilli P (1994) Interaction of Grb2 via its Src homology 3 domains with synaptic proteins including synapsin I. Proc Natl Acad Sci U S A 91(14):6486–6490CrossRefPubMedGoogle Scholar
  34. Moshe SL (1993) Seizures in the developing brain [J]. Neurology 43(11 Suppl 5):S3–S7PubMedGoogle Scholar
  35. Muramatsu R, Ikegaya Y, Matsuki N et al (2008) Early-life status epilepticus induces ectopic granule cells in adult mice dentate gyrus [J]. Exp Neurol 211(2):503–510CrossRefPubMedGoogle Scholar
  36. North BJ, Marshall BL, Borra MT, Denu JM, Verdin E (2003) The human Sir2 ortholog, SIRT2, is an NADþ-dependent tubulin deacetylase. Mol Cell 11(2):437–444CrossRefPubMedGoogle Scholar
  37. Oku T, Kaneko Y, Murofushi K, Seyama Y, Toyoshima S, Tsuji T (2008) Phorbol ester-dependent phosphorylation regulates the association of p57/coronin-1 with the actin cytoskeleton. J Biol Chem 283(43):28918–28925CrossRefPubMedGoogle Scholar
  38. Ouyang Y, Yang XF, Hu XY, Erbayat-Altay E, Zeng LH, Lee JM, Wong M (2007) Hippocampal seizures cause depolymerization of filamentous actin in neurons independent of acute morphological changes. Brain Res 1143:238–246CrossRefPubMedGoogle Scholar
  39. Sarac S, Afzal S, Broholm H, Madsen FF, Ploug T, Laursen H (2009) EAAT-1 and EAAT-2 in temporal lobe and hippocampus in intractable temporal lobe epilepsy. APMIS 117(4):291–301CrossRefPubMedGoogle Scholar
  40. Schuller E, Gulesserian T, Seidl R, Cairns N, Lubec G (2001) Brain t-complex polypeptide 1 (TCP-1) related to its natural substrate beta1 tubulin is decreased in Alzheimer’s disease. Life Sci 69(3):263–270CrossRefPubMedGoogle Scholar
  41. Schwartzkroin PA (1986) Hippocampal slices in experimental and human epilepsy. Adv Neurol 44:991–1010PubMedGoogle Scholar
  42. Shim KS, Lubec G (2002) Drebrin, a dendritic spine protein, is manifold decreased in brains of patients with Alzheimer’s disease and Down syndrome. Neurosci Lett 324(3):209–212CrossRefPubMedGoogle Scholar
  43. Terada S, Tsujimoto T, Takei Y, Takahashi T, Hirokawa N (1999) Impairment of inhibitory synaptic transmission in mice lacking synapsin I. J Cell Biol 145(5):1039–1048CrossRefPubMedGoogle Scholar
  44. Timofeev I, Bazhenov M, Avramescu S, Nita DA (2009) Posttraumatic epilepsy: the roles of synaptic plasticity. Neuroscientist (Epub ahead of print)Google Scholar
  45. Ueda S, Masutani H, Nakamura H, Tanaka T, Ueno M, Yodoi J (2002) Redox control of cell death. Antioxid Redox Signal 4(3):405–414CrossRefPubMedGoogle Scholar
  46. Weitzdoerfer R, Fountoulakis M, Lubec G (2002) Reduction of actin-related protein complex in fetal Down syndrome brain. Biochem Biophys Res Commun 293(2):836–841CrossRefPubMedGoogle Scholar
  47. Williams PA, Dou P, Dudek FE (2004) Epilepsy and synaptic reorganization in a perinatal rat model of hypoxia–ischemia. Epilepsia 45(10):1210–1218CrossRefPubMedGoogle Scholar
  48. Witke W, Podtelejnikov AV, Di Nardo A, Sutherland JD, Gurniak CB, Dotti C, Mann M (1998) In mouse brain profilin I and profilin II associate with regulators of the endocytic pathway and actin assembly. EMBO J 17(4):967–976CrossRefPubMedGoogle Scholar
  49. Wong M (2008) Stabilizing dendritic structure as a novel therapeutic approach for epilepsy. Expert Rev Neurother 8(6):907–915CrossRefPubMedGoogle Scholar
  50. Wu LW, Peng J, Wei CP, Yin F (2009) Preliminary explorations of differentially expressed proteins in hippocampus of MTLE rats following treatment with valproate, some findings from comparative proteomics. 13th Asian Pacific Congress of Pediatrics. Shanghai. OR710Google Scholar
  51. Xi JH, Bai F, McGaha R, Andley UP (2006) Alpha-crystallin expression affects microtubule assembly and prevents their aggregation. FASEB J 20(7):846–857CrossRefPubMedGoogle Scholar
  52. Yamagata Y (2003) New aspects of neurotransmitter release and exocytosis: dynamic and differential regulation of synapsin I phosphorylation by acute neuronal excitation in vivo. J Pharmacol Sci 93:22–29CrossRefPubMedGoogle Scholar
  53. Yang JW, Czech T, Felizardio M, Baumgartner C, Lubec G (2006) Aberrant expression of cytoskeleton proteins in hippocampus from patients with mesial temporal lobe epilepsy. Amino Acids 30(4):477–493CrossRefPubMedGoogle Scholar
  54. Yoo BC, Vlkolinsky R, Engidawork E, Cairns N, Fountoulakis M, Lubec G (2001) Differential expression of molecular chaperones in brain of patients with Down syndrome. Electrophoresis 22(6):1233–1241CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Liwen Wu
    • 1
  • Jing Peng
    • 1
  • Chaoping Wei
    • 1
  • Gu Liu
    • 1
  • Guoli Wang
    • 1
  • Kongzhao Li
    • 1
  • Fei Yin
    • 1
  1. 1.Department of PediatricsXiangya Hospital, Central South UniversityChangshaPeople’s Republic of China

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