Journal of Molecular Neuroscience

, Volume 67, Issue 3, pp 477–483 | Cite as

The Possible Role of Nitric Oxide Pathway in Pentylenetetrazole Preconditioning Against Seizure in Mice

  • Hedyeh Faghir-Ghanesefat
  • Hedieh Keshavarz-Bahaghighat
  • Nazanin Rajai
  • Tahmineh Mokhtari
  • Erfan Bahramnejad
  • Soheil Kazemi Roodsari
  • Ahmad Reza DehpourEmail author


Preconditioning is defined as an induction of adaptive response in organs against lethal stimulation provoked by subsequent mild sublethal stress. Several chemical agents have been demonstrated to cause brain tolerance through preconditioning. The aim of the present study is to test the hypothesis that preconditioning with pentylenetetrazole (PTZ) may have protective effect against seizure induced by i.v. infusion of PTZ. Mice were preconditioned by low-dose administration of PTZ (25 mg/kg) for 5 consecutive days, and the threshold of seizure elicited by i.v. infusion of PTZ was measured. To investigate the possible role of nitric oxide, NOS inhibitor enzymes, including L-NG-nitro-L-arginine methyl ester hydrochloride (L-NAME) (10 mg/kg), aminoguanidine (AG) (50 mg/kg), 7-nitroindazole (7-NI) (15 mg/kg), and L-arginine (L-arg) (60 mg/kg), were administered concomitantly with PTZ in both acute and chronic phases. Determination of seizure threshold revealed significant enhancement after preconditioning with low dose of PTZ. While the protective effect of PTZ preconditioning was enhanced after the administration of L-arg, it was reversed following administration of L-NAME and 7NI, suggesting the involvement of nitric oxide pathway as an underlying mechanism of PTZ-induced preconditioning. Preconditioning with PTZ led to brain tolerance and adaptive response in animal model of PTZ-induced seizure. This effect is in part due to the involvement of nitric oxide pathway.


Pentylenetetrazole Preconditioning Nitric oxide Seizure Mice 


Funding Statement

This study was funded and supported by Experimental Medicine Research Center, Tehran University of Medical Sciences (, grant no. 97-01-158-37318 to AD, and a grant (96002757) from Iran National Science Foundation (INSF).

Compliance with Ethical Standards

All applicable international and institutional guidelines for the care and use of animals were followed.

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. Amini E, Rezaei M, Ibrahim NM, Golpich M, Ghasemi R, Mohamed Z, Raymond AA, Dargahi L, Ahmadiani A (2015) A molecular approach to epilepsy management: from current therapeutic methods to preconditioning efforts. Mol Neurobiol 52(1):492–513Google Scholar
  2. Barone FC, White RF, Spera PA, Ellison J, Currie RW, Wang X, Feuerstein GZ (1998) Ischemic preconditioning and brain tolerance: temporal histological and functional outcomes, protein synthesis requirement, and interleukin-1 receptor antagonist and early gene expression. Stroke 29(9):1937–1951Google Scholar
  3. Bolli R (1996) The early and late phases of preconditioning against myocardial stunning and the essential role of oxyradicals in the late phase: an overview. Basic Res Cardiol 91(1):57–63Google Scholar
  4. Bredt DS, Hwang PM, Snyder SH (1990) Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature 347(6295):768–770Google Scholar
  5. Buisson A, Lakhmeche N, Verrecchia C, Plotkine M, Boulu R (1993) Nitric oxide: an endogenous anticonvulsant substance. Neuroreport 4(4):444–446Google Scholar
  6. Cocito L, Favale E, Reni L (1982) Epileptic seizures in cerebral arterial occlusive disease. Stroke 13(2):189–195Google Scholar
  7. Dahl NA, Balfour WM (1964) Prolonged anoxic survival due to anoxia pre-exposure: brain ATP, lactate, and pyruvate. Am J Phys 207(2):452–456Google Scholar
  8. de Araújo Herculano B, Vandresen-Filho S, Martins WC, Boeck CR, Tasca CI (2011) NMDA preconditioning protects against quinolinic acid-induced seizures via PKA, PI3K and MAPK/ERK signaling pathways. Behav Brain Res 219(1):92–97Google Scholar
  9. De Sarro G, Palma E, Costa N, Rosario M, Gratteri S, De Sarro A, Rotiroti D (2000) Effects of compounds acting on GABAB receptors in the pentylenetetrazole kindling model of epilepsy in mice. Neuropharmacology 39(11):2147–2161Google Scholar
  10. Denninger JW, Marletta MA (1999) Guanylate cyclase and the· NO/cGMP signaling pathway. Biochim Biophys Acta 1411(2–3):334–350Google Scholar
  11. Dirnagl U, Simon RP, Hallenbeck JM (2003) Ischemic tolerance and endogenous neuroprotection. Trends Neurosci 26(5):248–254Google Scholar
  12. Dmowska M, Cybulska R, Schoenborn R, Piersiak T, Jaworska-Adamu J, Gawron A (2010) Behavioural and histological effects of preconditioning with lipopolysaccharide in epileptic rats. Neurochem Res 35(2):262–272Google Scholar
  13. Gidday JM, Fitzgibbons JC, Shah AR, Park TS (1994) Neuroprotection from ischemic brain injury by hypoxic preconditioning in the neonatal rat. Neurosci Lett 168(1):221–224Google Scholar
  14. Gidday JM, Shah AR, Maceren RG, Wang Q, Pelligrino DA, Holtzman DM, Park T (1999) Nitric oxide mediates cerebral ischemic tolerance in a neonatal rat model of hypoxic preconditioning. J Cereb Blood Flow Metab 19(3):331–340Google Scholar
  15. Gorter JA, Pereira PMG, Van Vliet EA, Aronica E, Da Silva FHL, Lucassen PJ (2003) Neuronal cell death in a rat model for mesial temporal lobe epilepsy is induced by the initial status epilepticus and not by later repeated spontaneous seizures. Epilepsia 44(5):647–658Google Scholar
  16. Gupta Y, Chaudhary G, Srivastava AK (2002) Protective effect of resveratrol against pentylenetetrazole-induced seizures and its modulation by an adenosinergic system. Pharmacology 65(3):170–174Google Scholar
  17. Gupta YK, Veerendra Kumar MH, Srivastava AK (2003) Effect of Centella asiatica on pentylenetetrazole-induced kindling, cognition and oxidative stress in rats. Pharmacol Biochem Behav 74(3):579–585Google Scholar
  18. Heurteaux C, Lauritzen I, Widmann C, Lazdunski M (1995) Essential role of adenosine, adenosine A1 receptors, and ATP-sensitive K+ channels in cerebral ischemic preconditioning. Proc Natl Acad Sci U S A 92(10):4666–4670Google Scholar
  19. Huang R-Q, Bell-Horner CL, Dibas MI, Covey DF, Drewe JA, Dillon GH (2001) Pentylenetetrazole-induced inhibition of recombinant γ-aminobutyric acid type a (GABAA) receptors: mechanism and site of action. J Pharmacol Exp Ther 298(3):986–995Google Scholar
  20. Jimenez-Mateos EM, Henshall DC (2009) Seizure preconditioning and epileptic tolerance: models and mechanisms. Int J Physiol Pathophysiol Pharmacol 1(2):180–191Google Scholar
  21. Kato H, Araki T, Itoyama Y, Kogure K, Kato K (1995) An immunohistochemical study of heat shock protein-27 in the hippocampus in a gerbil model of cerebral ischemia and ischemic tolerance. Neuroscience 68(1):65–71Google Scholar
  22. Kitagawa K, Yagita Y, Sasaki T, Sugiura S, Omura-Matsuoka E, Mabuchi T, Matsushita K, Hori M (2005) Chronic mild reduction of cerebral perfusion pressure induces ischemic tolerance in focal cerebral ischemia. Stroke 36(10):2270–2274Google Scholar
  23. Kume M, Yamamoto Y, Saad S, Gomi T, Kimoto S, Shimabukuro T, Yagi T, Nakagami M, Takada Y, Morimoto T, Yamaoka Y (1996) Ischemic preconditioning of the liver in rats: implications of heat shock protein induction to increase tolerance of ischemia-reperfusion injury. J Lab Clin Med 128(3):251–258Google Scholar
  24. Maulik N, Engelman RM, Wei Z, Liu X, Rousou JA, Flack JE, Deaton DW, Das DK (1995) Drug-induced heat-shock preconditioning improves postischemic ventricular recovery after cardiopulmonary bypass. Circulation 92(9):381–388Google Scholar
  25. McCrohan CR, Gillette R (1988) Enhancement of cyclic AMP-dependent sodium current by the convulsant drug pentylenetetrazol. Brain Res 452(1–2):21–27Google Scholar
  26. Meller R, Cameron JA, Torrey DJ, Clayton CE, Ordonez AN, Henshall DC, Minami M, Schindler CK, Saugstad JA, Simon RP (2006) Rapid degradation of Bim by the ubiquitin-proteasome pathway mediates short-term ischemic tolerance in cultured neurons. J Biol Chem 281(11):7429–7436Google Scholar
  27. Meller R, Thompson SJ, Lusardi TA, Ordonez AN, Ashley MD, Jessick V, Wang W, Torrey DJ, Henshall DC, Gafken PR (2008) Ubiquitin–proteasome-mediated synaptic reorganization: a novel mechanism underlying rapid ischemic tolerance. J Neurosci 28(1):50–59Google Scholar
  28. Nakata N, Kato H, Kogure K (1994) Ischemic tolerance and extracellular amino acid concentrations in gerbil hippocampus measured by intracerebral microdialysis. Brain Res Bull 35(3):247–251Google Scholar
  29. National Research Council Committee for the Update of the Guide for the, C., Use of Laboratory, A (2011) The National Academies Collection: Reports funded by National Institutes of Health, in: th (Ed.) Guide for the Care and Use of Laboratory Animals. National Academies Press (US) National Academy of Sciences, Washington (DC)Google Scholar
  30. Neely CF, Keith IM (1995) A1 adenosine receptor antagonists block ischemia-reperfusion injury of the lung. Am J Physiol Lung Cell Mol Physiol 268(6):L1036–L1046Google Scholar
  31. Osonoe K, Mori N, Suzuki K, Osonoe M (1994) Antiepileptic effects of inhibitors of nitric oxide synthase examined in pentylenetetrazol-induced seizures in rats. Brain Res 663(2):338–340Google Scholar
  32. Rauca C, Rüthrich H-L (1995) Moderate hypoxia reduces pentylenetetrazol-induced seizures. Naunyn Schmiedeberg's Arch Pharmacol 351(3):261–267Google Scholar
  33. Rejdak R, Rejdak K, Sieklucka-Dziuba M, Stelmasiak Z, Grieb P (2001) Brain tolerance and preconditioning. Pol J Pharmacol 53(1):73–80Google Scholar
  34. Rubaj A, Gustaw K, Zgodziński W, Kleinrok Z, Sieklucka-Dziuba M (2000) The role of opioid receptors in hypoxic preconditioning against seizures in brain. Pharmacol Biochem Behav 67(1):65–70Google Scholar
  35. Saffen DW, Cole AJ, Worley PF, Christy BA, Ryder K, Baraban JM (1988) Convulsant-induced increase in transcription factor messenger RNAs in rat brain. Proc Natl Acad Sci U S A 85(20):7795–7799Google Scholar
  36. Sieklucka M, Bortolotto Z, Heim C, Block F, Sontag K (1991) Decreased susceptibility to seizures induced by bicuculline after transient bilateral clamping of the carotid arteries in rats. J Neural Transm Gen Sect 83(1–2):127–137Google Scholar
  37. Sieklucka M, Heim C, Block F, Sontag K-H (1992) Transient reduction of cerebral blood flow leads to longlasting increase in GABA content in vulnerable structures and decreased susceptibility to bicuculline induced seizures. J Neural Transm Gen Sect 88(2):87–94Google Scholar
  38. Tsikas D (2007) Analysis of nitrite and nitrate in biological fluids by assays based on the Griess reaction: appraisal of the Griess reaction in the L-arginine/nitric oxide area of research. J Chromatogr B Anal Technol Biomed Life Sci 851(1–2):51–70Google Scholar
  39. Vasconcelos AP, Gizard F, Marescaux C, Nehlig A (2000) Role of nitric oxide in Pentylenetetrazol-induced seizures: age-dependent effects in the immature rat. Epilepsia 41(4):363–371Google Scholar
  40. Yahyavi-Firouz-Abadi N, Tahsili-Fahadan P, Riazi K, Ghahremani MH, Dehpour AR (2006) Involvement of nitric oxide pathway in the acute anticonvulsant effect of melatonin in mice. Epilepsy Res 68(2):103–113Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Hedyeh Faghir-Ghanesefat
    • 1
    • 2
  • Hedieh Keshavarz-Bahaghighat
    • 1
    • 2
  • Nazanin Rajai
    • 1
    • 2
  • Tahmineh Mokhtari
    • 3
    • 4
  • Erfan Bahramnejad
    • 1
    • 2
  • Soheil Kazemi Roodsari
    • 1
    • 2
  • Ahmad Reza Dehpour
    • 1
    • 2
    Email author
  1. 1.Department of Pharmacology, School of MedicineTehran University of Medical SciencesTehranIran
  2. 2.Experimental Medicine Research CenterTehran University of Medical SciencesTehranIran
  3. 3.Nervous System Stem Cells Research CenterSemnan University of Medical SciencesSemnanIran
  4. 4.Department of Anatomy, School of MedicineSemnan University of Medical SciencesSemnanIran

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