Українська English

ISSN 2522-9028 (Print)
ISSN 2522-9036 (Online)
DOI: https://doi.org/10.15407/fz

Fiziologichnyi Zhurnal

is a scientific journal issued by the

Bogomoletz Institute of Physiology
National Academy of Sciences of Ukraine

Editor-in-chief: V.F. Sagach

The journal was founded in 1955 as
1955 – 1977 "Fiziolohichnyi zhurnal" (ISSN 0015 – 3311)
1978 – 1993 "Fiziologicheskii zhurnal" (ISSN 0201 – 8489)
1994 – 2016 "Fiziolohichnyi zhurnal" (ISSN 0201 – 8489)
2017 – "Fiziolohichnyi zhurnal" (ISSN 2522-9028)

Fiziol. Zh. 2021; 67(5): 3-10


THE EFFECT OF THROMBIN IN THE SERUM-ADAPTED IONIC ENVIRONMENT ON THE INDUCTION OF EPILEPTIFORM FIRING ACTIVITY OF HIPPOCAMPAL CULTURED NEURONS

M.S. Shypshyna, A.V. Savotchenko, K.I. Kuznetsov, M.S. Veselovsky

    O.O. Bogomolets Institute of Physiology of the NAS of Ukraine, Kyiv, Ukraine
DOI: https://doi.org/10.15407/fz67.05.003

Abstract

The mechanisms of epileptiform neuronal activity develop- ment under blood-brain barrier (BBB) dysfunction remains relevant in modern psychoneurology. In the present work we mimic some effects of BBB disruption in the culture of hip- pocampal neurons to examined the effect of serum-adapted ionic environment on the impulse activity of hippocampal neurons and the role of serum protein thrombin in induction of epileptiform neuronal activity. Using the whole-cell patch- clamp method under current-clamp mode we analyzed the spontaneous action potentials (AP) in the single hippocampal neurons. The changing of ionic extracellular neuronal environ- ment to such serum-adapted contributed to the development of epileptiform tonic activity of cultured hippocampal neurons and led to increase the average APs frequency by 65.1 ± 17.9% (n = 5) in neurons with spontaneous firing activity (FA) and to occurrence of tonic electrical activity (1.65 ± 0.4 s-1) in neurons without firing activity. Glutamate NMDA receptors significantly contribute to epileptiform tonic activity formation in neurons with FA, while their role in tonic activity providing in neurons without FA was insignificant. Thrombin (5 U/ml) in the serum-adapted ionic solution significantly enhanced of epileptiform activity in neurons with and without spontaneous FA: APs frequency increased in these neuronal groups by 117.3 ± 25.6% (n = 3) and by 61.8 ± 11.5% (n = 3), respective- ly, compared with that in the serum-adapted ionic solution only. Blockade of thrombin protease activated receptor 1 (PAR-1) by application of SCH 79797 (10 μm) canceled the thrombin’s effect in neurons without spontaneous FA, and significantly reduced such in neurons with FA. Therefore, the change of ionic extracellular neuronal environment to serum-adapted stimulates the occurrence of epileptiform activity in hippo- campal neurons, that is apparently associated with NMDA- receptors activation in neurons with FA. The proepileptiform action of thrombin was mostly mediated by PAR-1 activation. Thrombin-dependent regulation of the hippocampal single neurons firing activity involves the mechanisms different from the modulation of glutamate NMDA receptors in these cells.

Keywords: Key words: blood-brain barrier; epilepsy; hippocampus; action potentials; NMDA receptors; thrombin; PAR-1

References

  1. Reinert M, Khaldi A, Zauner A, Doppenberg E, Choi S, Bullock R. High extracellular potassium and its correlates after severe head injury: relationship to high intracranial pressure. Neurosurg Focus. 2000;8(1):1-8. CrossRef PubMed
  2. Zauner A, Bullock R, Kuta AJ, Woodward J, Young HF. Glutamate release and cerebral blood flow after severe human head injury. Acta Neurochir Suppl. 1996;67:40-4. CrossRef PubMed
  3. Luo BH, Carman CV, Springer TA. Structural basis of integrin regulation and signaling. Annu Rev Immunol. 2007;25:619-47. CrossRef PubMed PubMedCentral
  4. Maggio N, Shavit E, Chapman J, Segal M. 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. 2008;28:732-6. CrossRef PubMed PubMedCentral
  5. Isaeva E, Hernan A, Isaev D, Holmes GL. Thrombin fa- cilitates seizures through activation of persistent sodium current. Ann Neurol. 2012;72,192-8. CrossRef PubMed PubMedCentral
  6. Lee CJ, Mannaioni G, Yuan H, Woo DH, Gingrich MB, Traynelis SF. Astrocytic control of synaptic NMDA receptors. J Physiol. 2007;581,1057-81. CrossRef PubMed PubMedCentral
  7. Han KS, Mannaioni G, Hamill CE, Lee J, Junge CE, Lee CJ. Activation of protease activated receptor 1 increases the excitability of the dentate granule neurons of hip- pocampus. Mol Brain. 2011;4:32. CrossRef PubMed PubMedCentral
  8. Oh SJ, Han KS, Park H, Woo DH, Kim HY, Traynelis SF. Protease activated receptor 1-induced glutamate release in cultured astrocytes is mediated by Bestrophin-1 channel but not by vesicular exocytosis. Mol Brain. 2012;5:38. CrossRef PubMed PubMedCentral
  9. Park H, Han KS, Oh SJ, Jo S, Woo J, Yoon BE. High gluta- mate permeability and distal localization of Best1 channel in CA1 hippocampal astrocyte. Mol Brain. 2013;6:54. CrossRef PubMed PubMedCentral
  10. Mizerna OP, Fedulova SA, Veselovsky NS. Temporal regularity of neurotransmitter release at single ter- minal in cultured hippocampal neurons. Fiziol Zh. 2010;56(1):118-26. CrossRef
  11. Davson H and Segal MB. Physiology of the CSF and blood-brain barriers. Boca Raton, Fla: CRC Press; 1996.
  12. Katzman R and Pappius HM. Brain electrolytes and fluid metabolism. Baltimore: Williams &Wilkins; 1973.
  13. Somjen GG and Miiller M. Potassium-induced enhance- ment of persistent inward current in hippocampal neurons in isolation and in tissue slices. Brain Res. 2000;885,102-10. CrossRef
  14. Stafstrom CE. Persistent sodium current and its role in epilepsy. Epilepsy Current. 2007 Jan-Feb;7(1):15-22. CrossRef PubMed PubMedCentral
  15. Pandis C, Sotiriou E, Kouvaras E, Asprodini E, Pa- patheodoropoulos C, Angelatou F. Differential expression of NMDA and AMPA receptor subunits in rat dorsal and ventral hippocampus. Neuroscience. 2006;140:163-75. CrossRef PubMed
  16. Papatheodoropoulos C, Moschovos C, Kostopoulos G. Greater contribution of N-methyl-d-aspartic acid recep- tors in ventral compared to dorsal hippocampal slices in the expression and long-term maintenance of epileptiform activity. Neuroscience. 2005;135:765-79. CrossRef PubMed
  17. Isaev D, Lushnikova I, Lunko O, Zapukhliak O, Maximyuk O, Romanov A, Skibo GG, Tian C, Holmes GL, Isaeva E. Contribution of protease-activated receptor 1 in status epilepticus-induced epileptogenesis. Neurobiol Dis. 2015 Jun;78:68-76. CrossRef PubMed PubMedCentral
  18. Maggio N, Cavaliere C, Papa M, Blatt I, Chapman J, Segal M. Thrombin regulation of synaptic transmission: impli- cations for seizure onset. Neurobiol Dis. 2013;50:171-8. CrossRef PubMed
  19. Rice AC, DeLorenzo RJ. NMDA receptor activation dur- ing status epilepticus is required for the development of epilepsy. Brain Res. 1998;782(1-2):240-7. CrossRef
  20. Hellier JL, White A, Williams PA, Dudek FE, Staley KJ. NMDA receptor-mediated long-term alterations in epileptiform activity in experimental chronic epilepsy. Neuropharmacology. 2009;56(2):414-21. CrossRef PubMed PubMedCentral
  21. Ben Shimon M, Lenz M, Ikenberg B, Becker D, Shavit Stein E, Chapman J, Tanne D, Pick CG, Blatt I, Neufeld M, Vlachos A, Maggio N. Thrombin regulation of syn- aptic transmission and plasticity: implications for health and disease. Front Cell Neurosci. 2015;9:151. CrossRef PubMed PubMedCentral
  22. Gingrich MB, Junge CE, Lyuboslavsky P, Traynelis SF. Potentiation of NMDA receptor function by the serine protease thrombin. J Neurosci. 2000;20:4582-95. CrossRef PubMed PubMedCentral
  23. Becker D, Ikenberg B, Schiener S, Maggio N, Vlachos A. NMDA-receptor inhibition restores Protease-Activated Receptor 1 (PAR1) mediated alterations in homeostatic synaptic plasticity of denervated mouse dentate granule cells. Neuropharmacology. 2014;86,212-8. CrossRef PubMed
  24. Vance KM, Rogers RC, Hermann GE. PAR1-activated astrocytes in the nucleus of the solitary tract stimulate adjacent neurons via NMDA receptors. J Neurosci. 2015;35:776-85. CrossRef PubMed PubMedCentral

© National Academy of Sciences of Ukraine, Bogomoletz Institute of Physiology, 2014-2022.