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Suppressive effect of Rho-kinase inhibitors Y-27632 and fasudil on spike-and-wave discharges in genetic absence epilepsy rats from Strasbourg (GAERS)

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Abstract

Rho/Rho-kinase (ROCK) signaling contributes to neuroinflammation, epileptogenesis, and seizures in convulsive-type epilepsies. However, this pathway has not been investigated in absence epilepsy. We investigated RhoA activity in genetic absence epilepsy rats from Strasburg (GAERS) and the effects of ROCK inhibitors Y-27632 and fasudil on spike-and-wave discharges (SWDs) of GAERS. ROCK level and activity were measured by Western blot analysis in the brain areas involved in absence seizures (i.e., cortex and thalamus) and hippocampus. Male GAERS were stereotaxically implanted with bilateral cortical electrodes for electroencephalogram (EEG) recordings and/or guide cannula into the right ventricle. ROCK inhibitors were administered by intraperitoneal injection (1–10 mg/kg for Y-27632 or fasudil) or intracerebroventricular injection (7–20 nmol/5 μl for Y-27632 or 10–100 nmol/5 μl for fasudil). EEG was recorded under freely moving conditions. Compared with Wistar rats, GAERS exhibited increased RhoA activity in the somatosensory cortex but not in the thalamus or hippocampus. The single systemic administration of Y-27632 and fasudil partially suppressed the duration and frequency of absence seizure, respectively. However, local brain administration caused a widespread suppressive effect on the total seizure duration, number of seizures, and the average individual seizure length. In summary, Rho/ROCK signaling may be involved in the pathophysiology of absence epilepsy. Furthermore, ROCK inhibitors can control the expression of absence seizure in GAERS, thus indicating that Y-27632 and fasudil have the potential to be used as novel anti-absence drugs.

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References

  • Akin D, Ravizza T, Maroso M, Carcak N, Eryigit T, Vanzulli I, Aker RG, Vezzani A, Onat FY (2011) IL-1β is induced in reactive astrocytes in the somatosensory cortex of rats with genetic absence epilepsy at the onset of spike-and-wave discharges, and contributes to their occurrence. Neurobiol Dis 44:259–269

    Article  CAS  Google Scholar 

  • Avanzini G, de Curtis M, Marescaux C, Panzica F, Spreafico R, Vergnes M (1992) Role of the thalamic reticular nucleus in the generation of rhythmic thalamo-cortical activities subserving spike and waves. J Neural Transm Suppl 35:85–95

    CAS  PubMed  Google Scholar 

  • Blumenfeld H (2005) Cellular and network mechanisms of spike-wave seizures. Epilepsia 46(Suppl 9):21–33

    Article  CAS  Google Scholar 

  • Borisoff JF, Chan CCM, Hiebert GW, Oschipok L, Robertson GS, Zamboni R, Steeves JD, Tetzlaff W (2003) Suppression of rho-kinase activity promotes axonal growth on inhibitory CNS substrates. Mol Cell Neurosci 22:405–416

    Article  CAS  Google Scholar 

  • Büyükafşar K, Yalçin I, Kurt AH, Tiftik RN, Şahan-Firat S, Aksu F (2006) Rho-kinase inhibitor, Y-27632, has an antinociceptive effect in mice. Eur J Pharmacol 541:49–52

    Article  Google Scholar 

  • Çayan S, Saylam B, Tiftik N, Ünal ND, Apa DD, Efesoy O, Çimen B, Bozlu M, Akbay E, Büyükafşar K (2014) Rho-kinase levels in testicular ischemia-reperfusion injury and effects of its inhibitor, Y-27632, on oxidative stress, spermatogenesis, and apoptosis. Urology 83:675.e13–675.e18

    Article  Google Scholar 

  • Chen J, Yin W, Tu Y, Wang S, Yan X, Chen Q, Zhang X, Han Y, Pi R (2017) L-F001, a novel multifunctional ROCK inhibitor, suppresses neuroinflammation in vitro and in vivo: involvement of NF-κB inhibition and Nrf2 pathway activation. Eur J Pharmacol 806:1–9

    Article  CAS  Google Scholar 

  • Citraro R, Leo A, Marra R, De Sarro G, Russo E (2015) Antiepileptogenic effects of the selective COX-2 inhibitor etoricoxib, on the development of spontaneous absence seizures in WAG/Rij rats. Brain Res Bull 113:1–7

    Article  CAS  Google Scholar 

  • Crunelli V, Leresche N (2002) Childhood absence epilepsy: genes, channels, neurons and networks. Nat Rev Neurosci 3:371–382

    Article  CAS  Google Scholar 

  • Dash PK, Orsi SA, Moody M, Moore AN (2004) A role for hippocampal Rho-ROCK pathway in long-term spatial memory. Biochem Biophys Res Commun 322:893–898

    Article  CAS  Google Scholar 

  • Depaulis A, David O, Charpier S (2016) The genetic absence epilepsy rat from Strasbourg as a model to decipher the neuronal and network mechanisms of generalized idiopathic epilepsies. J Neurosci Methods 260:159–174

    Article  Google Scholar 

  • Dey A, Kang X, Qiu JG, Du YF, Jiang JX (2016) Anti-inflammatory small molecules to treat seizures and epilepsy: from bench to bedside. Trends Pharmacol Sci 37:463–484

    Article  CAS  Google Scholar 

  • Ding J, Li QY, Wang X, Sun CH, Lu CZ, Xiao BG (2010) Fasudil protects hippocampal neurons against hypoxia-reoxygenation injury by suppressing microglial inflammatory responses in mice. J Neurochem 114:1619–1629

    Article  CAS  Google Scholar 

  • Dubreuil CI, Marklund N, Deschamps K, McIntosh TK, McKerracher L (2006) Activation of Rho after traumatic brain injury and seizure in rats. Exp Neurol 198:361–369

    Article  CAS  Google Scholar 

  • Fukuta T, Asai T, Sato A, Namba M, Yanagida Y, Kikuchi T, Koide H, Shimizu K, Oku N (2016) Neuroprotection against cerebral ischemia/reperfusion injury by intravenous administration of liposomal fasudil. Int J Pharm 506:129–137

    Article  CAS  Google Scholar 

  • Gisselsson L, Toresson H, Ruscher K, Wieloch T (2010) Rho kinase inhibition protects CA1 cells in organotypic hippocampal slices during in vitro ischemia. Brain Res 1316:92–100

    Article  CAS  Google Scholar 

  • He Y, Xu H, Liang L, Zhan Z, Yang X, Yu X, Ye Y, Sun L (2008) Antiinflammatory effect of Rho kinase blockade via inhibition of NF-kappaB activation in rheumatoid arthritis. Arthritis Rheum 58:3366–3376

    Article  CAS  Google Scholar 

  • Henderson BW, Gentry EG, Rush T, Troncoso JC, Thambisetty M, Montine TJ, Herskowitz JH (2016) Rho-associated protein kinase 1 (ROCK1) is increased in Alzheimer’s disease and ROCK1 depletion reduces amyloid-β levels in brain. J Neurochem 138:525–531

    Article  CAS  Google Scholar 

  • Holmes MD, Brown M, Tucker DM (2004) Are “generalized” seizures truly generalized? Evidence of localized mesial frontal and frontopolar discharges in absence. Epilepsia 45:1568–1579

    Article  Google Scholar 

  • Inan S, Büyükafşar K (2008) Antiepileptic effects of two Rho-kinase inhibitors, Y-27632 and fasudil, in mice. Br J Pharmacol 155:44–51

    Article  CAS  Google Scholar 

  • Jeon BT, Jeong EA, Park SY, Son H, Shin HJ, Lee DH, Kim HJ, Kang SS, Cho GJ, Choi WS, Roh GS (2013) The Rho-kinase (ROCK) inhibitor Y-27632 protects against excitotoxicity-induced neuronal death in vivo and in vitro. Neurotox Res 23:238–248

    Article  CAS  Google Scholar 

  • Kennard JTT, Barmanray R, Sampurno S, Ozturk E, Reid CA, Paradiso L, D’Abaco GM, Kaye AH, Foote SJ, O’Brien TJ, Powell KL (2011) Stargazin and AMPA receptor membrane expression is increased in the somatosensory cortex of genetic absence epilepsy rats from Strasbourg. Neurobiol Dis 42:48–54

    Article  CAS  Google Scholar 

  • Kitaoka Y, Kitaoka Y, Kumai T, Lam TT, Kuribayashi K, Isenoumi K, Munemasa Y, Motoki M, Kobayashi S, Ueno S (2004) Involvement of RhoA and possible neuroprotective effect of fasudil, a rho kinase inhibitor, in NMDA-induced neurotoxicity in the rat retina. Brain Res 1018:111–118

    Article  CAS  Google Scholar 

  • Klein JP, Khera DS, Nersesyan H, Kimchi EY, Waxman SG, Blumenfeld H (2004) Dysregulation of sodium channel expression in cortical neurons in a rodent model of absence epilepsy. Brain Res 1000:102–109

    Article  CAS  Google Scholar 

  • Köksel O, Yildirim Ç, Tiftik RN, Kubat H, Tamer L, Cinel L, Kaplan MB, Deǧirmenci U, Özdülger A, Büyükafşar K (2005) Rho-kinase (ROCK-1 and ROCK-2) upregulation in oleic acid-induced lung injury and its restoration by Y-27632. Eur J Pharmacol 510:135–142

    Article  Google Scholar 

  • Kourdougli N, Varpula S, Chazal G, Rivera C (2015) Detrimental effect of post status epilepticus treatment with ROCK inhibitor Y-27632 in a pilocarpine model of temporal lobe epilepsy. Front Cell Neurosci 9:413

    Article  Google Scholar 

  • Kovács Z, Dobolyi A, Juhász G, Kékesi KA (2014) Lipopolysaccharide induced increase in seizure activity in two animal models of absence epilepsy WAG/Rij and GAERS rats and Long Evans rats. Brain Res Bull 104:7–18

    Article  Google Scholar 

  • Li M, Huang Y, Ma AAK, Lin E, Diamond MI (2009) Y-27632 improves rotarod performance and reduces huntingtin levels in R6/2 mice. Neurobiol Dis 36:413–420

    Article  CAS  Google Scholar 

  • Li YH, Yu JZ, Xin YL, Feng L, Chai Z, Liu JC, Zhang HZ, Zhang GX, Xiao BG, Ma CG (2015) Protective effect of a novel Rho kinase inhibitor WAR-5 in experimental autoimmune encephalomyelitis by modulating inflammatory response and neurotrophic factors. Exp Mol Pathol 99:220–228

    Article  CAS  Google Scholar 

  • Marescaux C, Vergnes M, Depaulis A (1992) Genetic absence epilepsy in rats from Strasbourg—a review. J Neural Transm Suppl 35:37–69

    CAS  PubMed  Google Scholar 

  • McCormick DA, Contreras D (2001) On the cellular and network bases of epileptic seizures. Annu Rev Physiol 63:815–846

    Article  CAS  Google Scholar 

  • Meeren HKM, Pijn JPM, van Luijtelaar ELJM, Coenen AML, Lopes da Silva FH (2002) Cortical focus drives widespread corticothalamic networks during spontaneous absence seizures in rats. J Neurosci 22:1480–1495

    Article  CAS  Google Scholar 

  • Nguyen T, Sherratt PJ, Nioi P, Yang CS, Pickett CB (2005) Nrf2 controls constitutive and inducible expression of ARE-driven genes through a dynamic pathway involving nucleocytoplasmic shuttling by Keap1. J Biol Chem 280:32485–32492

    Article  CAS  Google Scholar 

  • Mueller BK, Mack H, Teusch N (2005) Rho kinase, a promising drug target for neurological disorders. Nat Rev Drug Discov 4:387–398

    Article  CAS  Google Scholar 

  • O’Kane EM, Stone TW, Morris BJ (2004) Increased long-term potentiation in the CA1 region of rat hippocampus via modulation of GTPase signalling or inhibition of rho kinase. Neuropharmacology 46:879–887

    Article  Google Scholar 

  • Panayiotopoulos CP, Michael M, Sanders S, Valeta T, Koutroumanidis M (2008) Benign childhood focal epilepsies: assessment of established and newly recognized syndromes. Brain 131:2264–2286

    Article  Google Scholar 

  • Paxinos G, Watson C (1997) The rat brain in stereotaxic coordinates, 3rd edn. Academic, San Diego

    Google Scholar 

  • Polack P-O, Guillemain I, Hu E, Deransart C, Depaulis A, Charpier S (2007) Deep layer somatosensory cortical neurons initiate spike-and-wave discharges in a genetic model of absence seizures. J Neurosci 27:6590–6599

    Article  CAS  Google Scholar 

  • Polack PO, Mahon S, Chavez M, Charpier S (2009) Inactivation of the somatosensory cortex prevents paroxysmal oscillations in cortical and related thalamic neurons in a genetic model of absence epilepsy. Cereb Cortex 19:2078–2091

    Article  Google Scholar 

  • Ramer LM, Borisoff JF, Ramer MS (2004) Rho-kinase inhibition enhances axonal plasticity and attenuates cold hyperalgesia after dorsal rhizotomy. J Neurosci 24:10796–10805

    Article  CAS  Google Scholar 

  • Roser AE, Tönges L, Lingor P (2017) Modulation of microglial activity by Rho-kinase (ROCK) inhibition as therapeutic strategy in Parkinson’s disease and amyotrophic lateral sclerosis. Front Aging Neurosci 9:94

    Article  Google Scholar 

  • Russo E, Andreozzi F, Iuliano R, Dattilo V, Procopio T, Fiume G, Mimmi S, Perrotti N, Citraro R, Sesti G, Constanti A, De Sarro G (2014) Early molecular and behavioral response to lipopolysaccharide in the WAG/Rij rat model of absence epilepsy and depressive-like behavior, involves interplay between AMPK, AKT/mTOR pathways and neuroinflammatory cytokine release. Brain Behav Immun 42:157–168

    Article  CAS  Google Scholar 

  • Sato M, Tani E, Fujikawa H, Kaibuchi K (2000) Involvement of Rho-kinase-mediated phosphorylation of myosin light chain in enhancement of cerebral vasospasm. Circ Res 87:195–200

    Article  CAS  Google Scholar 

  • Segain JP, De la Blétière DR, Sauzeau V, Bourreille A, Hilaret G, Cario-Toumaniantz C, Pacaud P, Galmiche JP, Loirand G (2003) Rho kinase blockade prevents inflammation via nuclear factor κB inhibition: evidence in Crohn’s disease and experimental colitis. Gastroenterology 124:1180–1187

    Article  CAS  Google Scholar 

  • Sitnikova E, Kulikova S, Birioukova L, Raevsky VV (2011) Cellular neuropathology of absence epilepsy in the neocortex: a population of glial cells rather than neurons is impaired in genetic rat model. Acta Neurobiol Exp (Wars) 71(2):263–8

    Google Scholar 

  • Snead OC (1995) Basic mechanisms of generalized absence seizures. Ann Neurol 37(2):146–157

    Article  Google Scholar 

  • Steriade M, Contreras D (1995) Relations between cortical and thalamic cellular events during transition from sleep patterns to paroxysmal activity. J Neurosci 15:623–642

    Article  CAS  Google Scholar 

  • Steriade M, Contreras D (1998) Spike-wave complexes and fast components of cortically generated seizures. I. Role of neocortex and thalamus. J Neurophysiol 80:1439–1455

    Article  CAS  Google Scholar 

  • Tatenhorst L, Eckermann K, Dambeck V, Fonseca-Ornelas L, Walle H, Lopes da Fonseca T, Koch JC, Becker S, Tönges L, Bähr M, Outeiro TF, Zweckstetter M, Lingor P (2016) Fasudil attenuates aggregation of α-synuclein in models of Parkinson’s disease. Acta Neuropathol Commun 4:39

    Article  Google Scholar 

  • Tenney JR, Duong TQ, King JA, Ferris CF (2004) fMRI of brain activation in a genetic rat model of absence seizures. Epilepsia 45:576–582

    Article  Google Scholar 

  • Tsushima H, Mori M, Fujiwara N, Moriyama A (2003) Pharmacological characteristics of bombesin receptor mediating hypothermia in the central nervous system of rats. Brain Res 969:88–94

    Article  CAS  Google Scholar 

  • Wu X, Xu XM (2016) RhoA/Rho kinase in spinal cord injury. Neural Regen Res 11:23–27

    Article  Google Scholar 

  • Yamashita K, Kotani Y, Nakajima Y, Shimazawa M, Yoshimura S, Nakashima S, Iwama T, Hara H (2007) Fasudil, a Rho kinase (ROCK) inhibitor, protects against ischemic neuronal damage in vitro and in vivo by acting directly on neurons. Brain Res 1154:215–224

    Article  CAS  Google Scholar 

  • Zhou Y, Su Y, Li B, Liu F, Ryder JW, Wu X, Gonzalez-DeWhitt PA, Gelfanova V, Hale JE, May PC, Paul SM, Ni B (2003) Nonsteroidal anti-inflammatory drugs can lower amyloidogenic Abeta42 by inhibiting Rho. Science 302:1215–1217

    Article  CAS  Google Scholar 

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Acknowledgments

This project was supported by the Scientific and Technological Council of Turkey (TÜBİTAK-113S221). The authors are indebted to Berrin Çetin, Burcu Öztaş, and Ömer Gökhan Akarsu for their help in the Western blot experiments.

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Authors and Affiliations

  1. Department of Pharmacology Faculty of Pharmacy, Istanbul University, Istanbul, Turkey

    Nihan Çarçak

  2. Department of Medical Pharmacology, Faculty of Medicine, Marmara University, Istanbul, Turkey

    Melis Yavuz, Tuğba Eryiğit Karamahmutoğlu & Filiz Yılmaz Onat

  3. Department of Pharmacology Faculty of Medicine, Sütçüimam University, Kahramanmaras, Turkey

    Akif Hakan Kurt

  4. Department of Pharmacology, Faculty of Medicine, Mersin University, Mersin, Turkey

    Meral Urhan Küçük & Kansu Büyükafsar

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  1. Nihan Çarçak
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Contributions

NÇ, FYO, and KB conceived and designed the experiments. NÇ, MY, TE, AHK conducted the experiments. KB, AHK, and MUK assisted with the Western blot experiments. NÇ, KB, AHK analyzed the data. NÇ, FYO, and KB wrote the manuscript. All authors read and approved the manuscript.

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Correspondence to Filiz Yılmaz Onat.

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Çarçak, N., Yavuz, M., Eryiğit Karamahmutoğlu, T. et al. Suppressive effect of Rho-kinase inhibitors Y-27632 and fasudil on spike-and-wave discharges in genetic absence epilepsy rats from Strasbourg (GAERS). Naunyn-Schmiedeberg's Arch Pharmacol 391, 1275–1283 (2018). https://doi.org/10.1007/s00210-018-1546-9

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