Neurochemical effects of sleep deprivation in the hippocampus of the pilocarpine-induced rat model of epilepsy

Document Type : Original Article


1 Zoology Department, Faculty of Science, Cairo University, Giza, Egypt

2 Biophysics Department, Faculty of Science, Cairo University, Giza, Egypt

3 Medical Physiology Department, Medical Division, National Research Center, Giza, Egypt


Objective(s): The present study aims to investigate the pathological mechanisms mediating the effect of paradoxical sleep deprivation (PSD) for 48 hr on the spontaneous recurrent seizures (SRS) stage of the pilocarpine rat model of temporal lobe epilepsy.
Materials and Methods: This was carried out through assessment of amino acid neurotransmitter levels, the main oxidative stress parameters, and the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in the hippocampus. The experimental animals were divided into 4 groups: control, epileptic, PSD, and epileptic+PSD groups.
Results: Data indicated that PSD in epileptic rats induced a significant decrease in GSH levels. TNF-α increased significantly in the PSD group and decreased significantly in both epileptic rats and epileptic rats deprived of paradoxical sleep. PSD induced a significant increase in glutamine, glutamate, and aspartate and a significant decrease in GABA. In epileptic rats and epileptic rats deprived of PS, a significant increase in aspartate and a significant decrease in GABA and taurine were recorded.  
Conclusion: The present data suggest that exposure to PSD for 48 hr did not worsen the alterations produced in the present epileptic model. However, epileptic, PSD, epileptic + PSD groups showed a state of hyperexcitability and oxidative stress. PSD may increase the susceptibility of animals to the development of epilepsy. 


1. Hesdorffer DC, Logroscino G, Benn EK, Katri N, Cascino G, Hauser WA. Estimating risk for developing epilepsy: a population-based study in Rochester, Minnesota. Neurology 2011; 76: 23–27.
2. Dalby NO, Mody I. The process of epileptogenesis: a pathophysiological approach. Curr Opin Neurol 2001;14: 187–192.
3. Júnior HV, de FrançaFonteles MM, Mendes de Freitas R. Acute seizure activity promotes lipid peroxidation, increased nitrite levels and adaptive pathways against oxidative stress in the frontal cortex and striatum. Oxid Med Cell Longev 2009; 2:130-137.
4. Vezzani A, Friedman A, Dingledine RJ. The role of inflammation in epileptogenesis. Neuropharmacology 2013; 69: 16–24.
5. Manni R, Terzaghi M.  Comorbidity between epilepsy and sleep disorders. Epilepsy Res 2010; 90: 171–177.
6. Kotagal P, Yardi N. The relationship between sleep and epilepsy. Semin Pediatr Neurol 2008; 15: 42–49.
7. Steinsbek KS, Berg-Nielsen TS, Wichstrom L. Sleep disorders in preschoolers: prevalence and comorbidity with psychiatric symptoms. J Dev Behav Pediatr 2013; 34: 633–641.
8. Jacoby A, Snape D, Lane S, Baker GA. Self-reported anxiety and sleep problems in people with epilepsy and their association with quality of life. Epilepsy Behav 2015; 43: 149–158.
9. Gopalakrishnan A, Ji LL, Cirelli C. Sleep deprivation and cellular responses to oxidative stress. Sleep 2004; 27: 27–35.
10. Zielinski MR, McKenna JT, McCarley RW. Functions and Mechanisms of Sleep. AIMS Neurosci 2016;3:67-104.
11. Besedovsky L, Lange T, Born J. Sleep and immune function. Eur J Physiol 2012; 463: 121–137.
12. Oyanedel CN, Kelemen E, Scheller J, Born J, Rose-John S. Peripheral and central blockade of interleukin-6 trans-signaling differentially affects sleep architecture. Brain Behav Immun 2015; 50: 178-185.
13. Sinha SR. Basic Mechanisms of Sleep and Epilepsy. J Clin Neurophysiol 2011; 28: 103–110.
14. Díaz-Negrillo A. Influence of sleep and sleep deprivation on ictal and interictal epileptiform activity. Epilepsy Res Treat 2013; 2013:492524.
15. Tyng CM, Amin HU, Saad MNK,  Malik AS. The influences of emotion on learning and memory. Front Psychol 2017; 8: 1454.
16. Toyoda I, Bower MR, Leyva F, Buckmaster PS. Early activation of ventral hippocampus and subiculum during spontaneous seizures in a rat model of temporal lobe epilepsy. J Neurosci 2016; 33: 11100–11115.
17. Yang RH, Hou XH, Xu XN, Zhang L, Shi JN, F Wang et al. Sleep deprivation impairs spatial learning and modifies the hippocampal theta rhythm in rats. Neuroscience 2011; 173: 116–123.
18. Turski WA, Cavalheiro EA, Schwarz M, Czuczwar SJ, Kleinrok Z, Turski L. Limbic seizures produced by pilocarpine in rats: behavioural, electroencephalographic and neuropathological study. Behav Brain Res 1983; 9: 315–335.
19. Williams MB, Jope RS. Protein synthesis inhibitors attenuate seizures induced inrats by lithium plus pilocarpine. Exp Neurol 1994;129: 169–173.
20. Cavalheiro EA, Leite JP, Bortolotto ZA, Turski WA, Ikonomidou C, Turski L. Long term effects of pilocarpine in rats: structural damage of the brain triggers kindling and spontaneous recurrent seizures. Epilepsia 1991; 32: 778–782.
21. Zager A, Andersen ML, Lima MM, Reksidler AB, Machado RB, Tufik S. Modulation of sickness behavior by sleep: the role of neurochemical and neuroinflammatory pathways in mice. Eur Neuropsychopharmacol 2009; 19: 589–602.
22. Màrquez FJ, Quesada AR, Sάnchez-Jiménez F, Múñez De Castro I. Determination of 27 dansyl amino acid derivatives in biological fluids by reversed-phase high performance liquid chromatography. J Chromatogr 1986; 380: 275–283.
23. Ruiz-Larrea MB, Leal AM, Liza M, Lacort M, de Groot H. Antioxidant effects of estradiol and 2-hydroxyestradiol on iron induced lipid peroxidation of rat liver microsomes, Steroids 1994; 59: 383–388.
24. Moshage H, Kok B, Huizenga JR. Nitrite and nitrate determination in plasma: a critical evaluation. Clin Chem 41; 1995; 892–896.
25. Ellman GL. Tissue sulfhydryl groups, Arch Biochem 1959; 82: 70–77.
 26.  Ellman GL, Courtney KD, Andres V, Feather-Stone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 1961; 7: 88–95.
27. Gorun V, Proinov I, Baltescu V, Balaban G, Barzu O. Modified Ellman procedure for assay of cholinesterase in crude enzymatic preparations. Anal Biochem 1978; 86: 324–326.
28. Mohammed HS, AboulEzz HS, Khadrawy YK, Noor NA. Neurochemical and electrophysiological changes induced by paradoxical sleep deprivation in rats. Behav Brain Res 2011; 225: 39-46.
29. Planagumà J, Leypoldt F, Mannara F, Gutiérrez-Cuesta J, Martín-García E, Aguilar E, et al. Human N-methyl D-aspartate receptor antibodies alter memory and behaviour in mice. Brain. 2015;138:94-109.
30. Dong X, Wang Y, Qin Z. Molecular mechanisms of excitotoxicity and their relevance to pathogenesis of neurodegenerative diseases. Acta Pharmacol Sin  2009; 30: 379–387.
31. Dash MB, Douglas CL, Vyazovskiy VV, Cirelli C, Tononi G. Long-term homeostasis of extracellular glutamate in the rat cerebral cortex across sleep and waking states. J Neurosci. 2009: 21;29:620-69.
32. Khadrawy YK, Nour NA, AboulEzz HS. Effect of oxidative stress induced by paradoxical sleep deprivation on the activities of Na+, K+-ATPase and acetylcholinesterase in the cortex and hippocampus of rat. Transl Res 2011; 157: 100–107.
33. Silvestri AJ. REM sleep deprivation affects extinction of cued but not contextual fear conditioning. Physiol Behav 2005; 84:343–349.
34. Pickering M, Cumiskey D, O’Connor JJ. Actions of TNF-alpha on glutamatergic synaptic transmission in the central nervous system. Exp Physiol 2005; 90: 663–670.
35. Hsu JC, Lee YS, Chang CN, Chuang HL, Ling EA, Lan CT. Sleep deprivation inhibits expression of NADPH-d and NOS while activating microglia and astroglia in the rat hippocampus. Cells Tissues Organs 2009; 173: 242-254.
36. Rosso IM, Crowley DJ, Silveri MM, Rauch SL, Jensen JE. Hippocampus Glutamate and N-Acetyl Aspartate Markers of Excitotoxic Neuronal Compromise in Posttraumatic Stress Disorder. Neuropsychopharmacology 2017;42:1698-1705.
37. Nadler JV. Aspartate release and signalling in the hippocampus. Neurochem Res 2011; 36:668-676.
38. Noor NA, Aboul Ezz HS, Faraag AR, Khadrawy YA. Evaluation of the antiepileptic effect of curcumin and Nigella sativa oil in the pilocarpine model of epilepsy in comparison with valproate. Epilepsy Behav 2012; 24: 199–206.
39. Sgadò P, Dunleavy M, Genovesi S, Provenzano G, Bozzi Y. The role of GABAergic system in neurodevelopmental disorders: a focus on autism and epilepsy. Int J Physiol Pathophysiol Pharmacol 2011; 3: 223–235.
40. El Idrissi A. Taurine increases mitochondrial buffering of calcium: Role in neuroprotection. Amino Acids 2008:34:321–328.
41. Pahuja M, Mehla J, Reeta KH, Joshi S, Gupta YK. Hydroalcoholic extract of Zizyphusjujuba ameliorates seizures, oxidative stress, and cognitive impairment in experimental models of epilepsy in rats. Epilepsy Behav 2011; 21: 356–363.
42. Simonie´ A,  Laginja J, Varljen J, Laginja J, Simonić A. Lithium plus pilocarpine induced status epilepticus—biochemical changes. Neurosci Res 2000; 36: 157–166
43. Aboul Ezz HS, Khadrawy YA,  Noor NA. The neuroprotective effect of curcumin and nigella sativa oil against oxidative stress in the pilocarpine model of epilepsy: A comparison with valproate. Neurochem Res 2011; 36: 2195–2204.
44. Freitas RM. Investigation of oxidative stress involvement in hippocampus in epilepsy model induced by pilocarpine. Neurosci Lett 2009; 462: 225–229.
45. Kovács R, Rabanus A, Otáhal J, Patzak A, Kardos J, Albus K, et al. Endogenous nitric oxide is a key promoting factor for initiation of seizure-like events in hippocampal and entorhinal cortex slices. J Neurosci 2009; 29: 8565–8577.
46. Cupello A, Mainardi P, Robello M, Thellung S. Effect of nitric oxide donors on GABA uptake by rat brain synaptosomes. Neurochem. Res 1997; 22: 1517-1521.
47. Yuhas Y, Weizman A,  Ashkenazi S. Bidirectional concentration-dependent effects of tumor necrosis factor alpha in Shigella dysenteriae-related seizures. Infect Immun 2003; 71: 2288–2291.
48. Rana A, Musto AE. The role of inflammation in the development of epilepsy. J Neuroinflammation 2018;15:144-155.
49.  Murillo-Rodriguez E, Arias-Carrion O, Zavala-Garcia A, Sarro-Ramirez A, Huitron-Resendiz S, Arankowsky-Sandoval G. Basic sleep mechanisms: an integrative review. Cent Nerv Syst Agents Med Chem 2012; 12(1):38–54.
50. Merino JJ, Arce C, Naddaf A, Bellver-Landete V, Oset-Gasque MJ, González MP. The nitric oxide donor SNAP-induced amino acid neurotransmitter release in cortical neurons. Effects of blockers of voltage-dependent sodium and calcium channels. PLoS One 2014;9:e90703.
51. Ramanathan L, Hu S, Frautschy SA, Siegel JM. Short-term total sleep deprivation in the rat increases antioxidant responses in multiple brain regions without impairing spontaneous alternation behavior. Behav Brain Res 2010;207:305-309.
52. Wyse AT, Stefanello FM, Chiarani F, Delwing D, Wannmacher CM, Wajner M. Arginine administration decreases cerebral cortex acetylcholinesterase and serum butyrylcholinesterase probably by oxidative stress induction. Neurochem Res 2004; 29: 385–389.
53. Tsakiris S, Angelogianni P, Schulpis KH, Stavridis JC. Protective effect of L-phenylalanine on rat brain acetylcholinesterase inhibition induced by free radicals. Clin Biochem 2000; 33: 103–106.