Journal of Molecular Neuroscience

, Volume 67, Issue 2, pp 227–234 | Cite as

Thrombin and the Protease-Activated Receptor-1 in Organophosphate-Induced Status Epilepticus

  • Valery GoldermanEmail author
  • Efrat Shavit-Stein
  • Orna Gera
  • Joab Chapman
  • Arik Eisenkraft
  • Nicola Maggio


Organophosphates (OP) are a major threat to the health of soldiers and civilians due to their use as chemical weapons in war and in terror attacks. Among the acute manifestations of OP poisoning, status epilepticus (SE) is bearing the highest potential for long-term damages. Current therapies do not prevent brain damage and seizure-related brain injuries in OP-exposed humans. Thrombin is a serine protease known to have a fundamental function in the clotting cascade. It is highly expressed in the brain where we have previously found that it regulates synaptic transmission and plasticity. In addition, we have found that an excess of thrombin in the brain leads to hyperexcitability and therefore seizures through a glutamate-dependent mechanism. In the current study, we carried out in vitro, ex vivo, and in vivo experiments in order to determine the role of thrombin and its receptor PAR-1 in paraoxon-induced SE. Elevated thrombin activity was found in the brain slices from mice that were treated (in vitro and in vivo) with paraoxon. Increased levels of PAR-1 and pERK proteins and decreased prothrombin mRNA were found in the brains of paraoxon-treated mice. Furthermore, ex vivo and in vivo electrophysiological experiments showed that exposure to paraoxon causes elevated electrical activity in CA1 and CA3 regions of the hippocampus. Moreover, a specific PAR-1 antagonist (SCH79797) reduced this activity. Altogether, these results reveal the importance of thrombin and PAR-1 in paraoxon poisoning. In addition, the results indicate that thrombin and PAR-1 may be a possible target for the treatment of paraoxon-induced status epilepticus.


Thrombin PAR-1 Status epilepticus Organophosphates Paraoxon 



The authors thank Raphael Rosenbaum for his contribution in editing and proofreading of this manuscript.


  1. Araque A, Martín ED, Perea G, Arellano JI, Buño W (2002) Synaptically released acetylcholine evokes Ca2+ elevations in astrocytes in hippocampal slices. J Neurosci 22(7):2443–2450CrossRefPubMedGoogle Scholar
  2. Bushi D, Chapman J, Katzav A, Shavit-Stein E, Molshatzki N, Maggio N, Tanne D (2013) Quantitative detection of thrombin activity in an ischemic stroke model. J Mol Neurosci 51(3):844–850. CrossRefPubMedGoogle Scholar
  3. Chapman J (2013) Coagulation in inflammatory diseases of the central nervous system. Semin Thromb Hemost 39(8):876–880. CrossRefPubMedGoogle Scholar
  4. Citron BA, Smirnova IV, Zoubine MN, Festoff BW (1997) Quantitative pcr analysis reveals novel expression of prothrombin mrna and regulation of its levels in developing mouse muscle. Thromb Res 87(3):303–313. CrossRefPubMedGoogle Scholar
  5. Eisenkraft A, Falk A, Finkelstein A (2013) The role of glutamate and the immune system in organophosphate-induced CNS damage. Neurotox Res 24:265–279. CrossRefPubMedGoogle Scholar
  6. Golderman V, Shavit-Stein E, Tamarin I, Rosman Y, Shrot S, Rosenberg N et al (2016) The organophosphate paraoxon and its antidote obidoxime inhibit thrombin activity and affect coagulation in vitro. PLoS One 11(9):e0163787. CrossRefPubMedGoogle Scholar
  7. Greig NH, Reale M, Tata AM (2013) New pharmacological approaches to the cholinergic system: an overview on muscarinic receptor ligands and cholinesterase inhibitors. Recent Pat CNS Drug Discov 8(2):123–141CrossRefPubMedGoogle Scholar
  8. Isaev D, Lushnikova I, Lunko O, Zapukhliak O, Maximyuk O, Romanov A, Isaeva E (2015) Contribution of protease-activated receptor 1 in status epilepticus-induced epileptogenesis. Neurobiol Dis 78:68–76. CrossRefPubMedGoogle Scholar
  9. Junge CE, Lee CJ, Hubbard KB, Zhang Z, Olson JJ, Hepler JR et al (2004) Protease-activated receptor-1 in human brain: localization and functional expression in astrocytes. Exp Neurol 188:94–103. CrossRefPubMedGoogle Scholar
  10. Maggio N, Shavit E, Chapman J, Segal M (2008) Thrombin induces long-term potentiation of reactivity to afferent stimulation and facilitates epileptic seizures in rat hippocampal slices: toward understanding the functional consequences of cerebrovascular insults. J Neurosci 28:732–736. CrossRefPubMedGoogle Scholar
  11. Maggio N, Blatt I, Vlachos A, Tanne D, Chapman J, Segal M (2013a) Treating seizures and epilepsy with anticoagulants? Front Cell Neurosci 7:19. CrossRefPubMedGoogle Scholar
  12. Maggio N, Cavaliere C, Papa M, Blatt I, Chapman J, Segal M (2013b) Thrombin regulation of synaptic transmission: implications for seizure onset. Neurobiol Dis 50:171–178. CrossRefPubMedGoogle Scholar
  13. Maggio N, Itsekson Z, Dominissini D, Blatt I, Amariglio N, Rechavi G, Chapman J (2013c) Thrombin regulation of synaptic plasticity: implications for physiology and pathology. Exp Neurol 247:595–604. CrossRefPubMedGoogle Scholar
  14. Markel G, Krivoy A, Rotman E, Schein O, Shrot S, Brosh-Nissimov T et al (2008) Medical management of toxicological mass casualty events. Isr Med Assoc J 10:761–766PubMedGoogle Scholar
  15. McDonough JH Jr, Shih TM (1997) Neuropharmacological mechanisms of nerve agent-induced seizure and neuropathology. Neurosci Biobehav Rev 21:559–579CrossRefPubMedGoogle Scholar
  16. Paxinos G, Franklin KBJ (2004) The mouse brain in stereotaxic coordinates, vol 2nd. Academic Press.
  17. Pompili E, Nori SL, Geloso MC, Guadagni E, Corvino V, Michetti F, Fumagalli L (2004) Trimethyltin-induced differential expression of PAR subtypes in reactive astrocytes of the rat hippocampus. Mol Brain Res 122(1):93–98. CrossRefPubMedGoogle Scholar
  18. Quistad GB, Casida JE (2000) Sensitivity of blood-clotting factors and digestive enzymes to inhibition by organophosphorus pesticides. J Biochem Mol Toxicol 14:51–56CrossRefPubMedGoogle Scholar
  19. Shavit E, Michaelson DM, Chapman J (2011) Anatomical localization of protease-activated receptor-1 and protease-mediated neuroglilal crosstalk on peri-synaptic astrocytic endfeet. J Neurochem 119(3):460–473. CrossRefPubMedGoogle Scholar
  20. Shrot S, Markel G, Dushnitsky T, Krivoy A (2009) The possible use of oximes as antidotal therapy in organophosphate-induced brain damage. Neurotoxicology 30:167–173. CrossRefPubMedGoogle Scholar
  21. Striggow F, Riek M, Breder J, Henrich-Noack P, Reymann KG, Reiser G (2000) The protease thrombin is an endogenous mediator of hippocampal neuroprotection against ischemia at low concentrations but causes degeneration at high concentrations. Proc Natl Acad Sci U S A 97(5):2264–2269. CrossRefPubMedGoogle Scholar
  22. Sweeney AM, Fleming KE, McCauley JP, Rodriguez MF, Martin ET, Sousa AA, Scimemi A (2017) PAR1 activation induces rapid changes in glutamate uptake and astrocyte morphology. Sci Rep 7(1):43606. CrossRefPubMedGoogle Scholar
  23. ​Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76(9):4350–4354Google Scholar
  24. Ubl J, Vöhringer C, Reiser G (1998) Co-existence of two types of [Ca2+]i-inducing protease-activated receptors (PAR-1 and PAR-2) in rat astrocytes and C6 glioma cells. Neuroscience 86(2):597–609. CrossRefPubMedGoogle Scholar
  25. Weissman BA, Raveh L (2008) Therapy against organophosphate poisoning: the importance of anticholinergic drugs with antiglutamatergic properties. Toxicol Appl Pharmacol 232:351–358. CrossRefPubMedGoogle Scholar
  26. Zhao P, Metcalf M, Bunnett NW (2014) Biased signaling of protease-activated receptors. Front Endocrinol 5.

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of NeurologyThe Chaim Sheba Medical CenterRamat GanIsrael
  2. 2.Joseph Sagol Neuroscience CenterSheba Medical CenterRamat GanIsrael
  3. 3.Department of Physical Therapy, Sackler Faculty of MedicineTel Aviv UniversityTel AvivIsrael
  4. 4.Department of Neurology, Sackler School of MedicineTel Aviv UniversityTel AvivIsrael
  5. 5.Robert and Martha Harden Chair in Mental and Neurological Diseases, Sackler Faculty of MedicineTel AvivIsrael
  6. 6.Institute for Research in Military MedicineThe Hebrew University of JerusalemJerusalemIsrael
  7. 7.Talpiot medical leadership programChaim Sheba Medical CenterRamat GanIsrael

Personalised recommendations