Cellular and Molecular Neurobiology

, Volume 31, Issue 3, pp 469–478

Effect of the PARP-1 Inhibitor PJ 34 on Excitotoxic Damage Evoked by Kainate on Rat Spinal Cord Organotypic Slices

Original Research

Abstract

Excitotoxicity triggered by over-activation of glutamate receptors is thought to be an early mechanism of extensive neuronal death with consequent loss of function following lesion of spinal networks. One important process responsible for excitotoxic death is ‘parthanatos’ caused by hyperactivation of poly(ADP-ribose) polymerase (PARP) enzyme 1. Using rat organotypic spinal slices as in vitro models, the present study enquired if 2-(dimethylamino)-N-(5,6-dihydro-6-oxophenanthridin-2yl)acetamide (PJ 34), a pharmacological inhibitor of PARP-1, could counteract the excitotoxic damage evoked by transient application (1 h) of kainate, a potent analogue of glutamate. Kainate induced dose-dependent (1 μM threshold) neuronal loss (without damage to astrocytes) detected 24 h later via a PARP-1 dependent process that had peaked at 4 h after washout kainate. All spinal regions (ventral, central and dorsal) were affected, even though the largest damage was found in the dorsal area. Whereas PJ 34 did not protect against a large concentration (100 μM) of kainate, it significantly inhibited neuronal losses evoked by 10 μM kainate as long as it was co-applied with this glutamate agonist. When the application of PJ 34 was delayed to the washout time, neuroprotection was weak and regionally restricted. These data suggest that kainate-induced parthanatos developed early and was prevented by PJ 34 only when it was co-applied together with excitotoxic stimulus. Our results highlight the difficulty to arrest parthanatos as a mechanism of spinal neuron death in view of its low threshold of activation by kainate, its widespread distribution, and relatively fast development.

Keywords

Organotypic culture Kainic acid Parthanatos Spinal cord injury Excitotoxicity 

References

  1. Abdelkarim GE, Gertz K, Harms C, Katchanov J, Dirnagl U, Szabo C, Endres M (2001) Protective effects of PJ34, a novel, potent inhibitor of poly(ADP-ribose) polymerase (PARP) in in vitro and in vivo models of stroke. Int J Mol Med 7:255–260PubMedGoogle Scholar
  2. Agrawal SG, Evans RH (1986) The primary afferent depolarizing action of kainate in the rat. Br J Pharmacol 87:345–355PubMedGoogle Scholar
  3. Agrawal SM, Lau L, Yong VW (2008) MMPs in the central nervous system: where the good guys go bad. Semin Cell Dev Biol 19:42–51CrossRefPubMedGoogle Scholar
  4. Andrabi SA, Dawson TM, Dawson VL (2008) Mitochondrial and nuclear cross talk in cell death: parthanatos. Ann N Y Acad Sci 1147:233–241CrossRefPubMedGoogle Scholar
  5. Besson VC (2009) Drug targets for traumatic brain injury from poly(ADP-ribose)polymerase pathway modulation. Br J Pharmacol 157:695–704CrossRefPubMedGoogle Scholar
  6. Calderó J, Brunet N, Tarabal O, Piedrafita L, Hereu M, Ayala V, Esquerda JE (2010) Lithium prevents excitotoxic cell death of motoneurons in organotypic slice cultures of spinal cord. Neuroscience 165:1353–1369CrossRefPubMedGoogle Scholar
  7. Casey PJ, Black JH, Szabo C, Frosch M, Albadawi H, Chen M, Cambria RP, Watkins MT (2005) Poly(adenosine diphosphate ribose) polymerase inhibition modulates spinal cord dysfunction after thoracoabdominal aortic ischemia-reperfusion. J Vasc Surg 41:99–107CrossRefPubMedGoogle Scholar
  8. Cater HL, Gitterman D, Davis SM, Benham CD, Morrison B III, Sundstrom LE (2007) Stretch-induced injury in organotypic hippocampal slice cultures reproduces in vivo post-traumatic neurodegeneration: role of glutamate receptors and voltage-dependent calcium channels. J Neurochem 101:434–447CrossRefPubMedGoogle Scholar
  9. Corse AM, Bilak MM, Bilak SR, Lehar M, Rothstein JD, Kuncl RW (1999) Preclinical testing of neuroprotective neurotrophic factors in a model of chronic motor neuron degeneration. Neurobiol Dis 6:335–346CrossRefPubMedGoogle Scholar
  10. Doble A (1999) The role of excitotoxicity in neurodegenerative disease: implications for therapy. Pharmacol Ther 81:163–221CrossRefPubMedGoogle Scholar
  11. Donato R (2003) Intracellular and extracellular roles of S100 proteins. Microsc Res Tech 60:540–551CrossRefPubMedGoogle Scholar
  12. Formentini L, Macchiarulo A, Cipriani G, Camaioni E, Rapizzi E, Pellicciari R, Moroni F, Chiarugi A (2009) Poly(ADP-ribose) catabolism triggers AMP-dependent mitochondrial energy failure. J Biol Chem 284:17668–17676CrossRefPubMedGoogle Scholar
  13. Fossati S, Cipriani G, Moroni F, Chiarugi A (2007) Neither energy collapse nor transcription underlie in vitro neurotoxicity of poly(ADP-ribose) polymerase hyper-activation. Neurochem Int 50:203–210CrossRefPubMedGoogle Scholar
  14. Giansanti V, Dona F, Tillhon M, Scovassi AI (2010) PARP inhibitors: new tools to protect from inflammation. Biochem Pharmacol 80:1869–1877CrossRefPubMedGoogle Scholar
  15. Goebel DJ, Winkler BS (2006) Blockade of PARP activity attenuates poly(ADP-ribosyl)ation but offers only partial neuroprotection against NMDA-induced cell death in the rat retina. J Neurochem 98:1732–1745CrossRefPubMedGoogle Scholar
  16. Kauppinen TM, Suh SW, Berman AE, Hamby AM, Swanson RA (2009) Inhibition of poly(ADP-ribose) polymerase suppresses inflammation and promotes recovery after ischemic injury. J Cereb Blood Flow Metab 29:820–829CrossRefPubMedGoogle Scholar
  17. Kristensen BW, Noraberg J, Zimmer J (2001) Comparison of excitotoxic profiles of ATPA, AMPA, KA and NMDA in organotypic hippocampal slice cultures. Brain Res 917:21–44CrossRefPubMedGoogle Scholar
  18. Kuzhandaivel A, Margaryan G, Nistri A, Mladinic M (2010a) Extensive glial apoptosis develops early after hypoxic-dysmetabolic insult to the neonatal rat spinal cord in vitro. Neuroscience 169:325–338CrossRefPubMedGoogle Scholar
  19. Kuzhandaivel A, Nistri A, Mladinic M (2010b) Kainate-mediated excitotoxicity induces neuronal death in the rat spinal cord in vitro via a PARP-1 dependent cell death pathway (Parthanatos). Cell Mol Neurobiol 30:1001–1012CrossRefPubMedGoogle Scholar
  20. Lau A, Tymianski M (2010) Glutamate receptors, neurotoxicity and neurodegeneration. Pflugers Arch 460:525–542CrossRefPubMedGoogle Scholar
  21. Maragakis NJ, Rao MS, Llado J, Wong V, Xue H, Pardo A, Herring J, Kerr D, Coccia C, Rothstein JD (2005) Glial restricted precursors protect against chronic glutamate neurotoxicity of motor neurons in vitro. Glia 50:145–159CrossRefPubMedGoogle Scholar
  22. Mazzone GL, Margaryan G, Kuzhandaivel A, Nasrabady SE, Mladinic M, Nistri A (2010) Kainate-induced delayed onset of excitotoxicity with functional loss unrelated to the extent of neuronal damage in the in vitro spinal cord. Neuroscience 168:451–462CrossRefPubMedGoogle Scholar
  23. Moroni F (2008) Poly(ADP-ribose)polymerase 1 (PARP-1) and postischemic brain damage. Curr Opin Pharmacol 8:96–103CrossRefPubMedGoogle Scholar
  24. Moroni F, Formentini L, Gerace E, Camaioni E, Pellegrini-Giampietro DE, Chiarugi A, Pellicciari R (2009) Selective PARP-2 inhibitors increase apoptosis in hippocampal slices but protect cortical cells in models of post-ischaemic brain damage. Br J Pharmacol 157:854–862CrossRefPubMedGoogle Scholar
  25. Nicolescu AC, Holt A, Kandasamy AD, Pacher P, Schulz R (2009) Inhibition of matrix metalloproteinase-2 by PARP inhibitors. Biochem Biophys Res Commun 387:646–650CrossRefPubMedGoogle Scholar
  26. Nistri A, Taccola G, Mladinic M, Margaryan G, Kuzhandaivel A (2010) Deconstructing locomotor networks with experimental injury to define their membership. Ann N Y Acad Sci 1198:242–251CrossRefPubMedGoogle Scholar
  27. Park E, Velumian AA, Fehlings MG (2004) The role of excitotoxicity in secondary mechanisms of spinal cord injury: a review with an emphasis on the implications for white matter degeneration. J Neurotrauma 21:754–774CrossRefPubMedGoogle Scholar
  28. Rowland JW, Hawryluk GW, Kwon B, Fehlings MG (2008) Current status of acute spinal cord injury pathophysiology and emerging therapies: promise on the horizon. Neurosurg Focus 25:E2CrossRefPubMedGoogle Scholar
  29. Sandhu SK, Yap TA, de Bono JS (2010) Poly(ADP-ribose) polymerase inhibitors in cancer treatment: a clinical perspective. Eur J Cancer 46:9–20CrossRefPubMedGoogle Scholar
  30. Sibilla S, Ballerini L (2009) GABAergic and glycinergic interneuron expression during spinal cord development: dynamic interplay between inhibition and excitation in the control of ventral network outputs. Prog Neurobiol 89:46–60CrossRefPubMedGoogle Scholar
  31. Sibilla S, Fabbro A, Grandolfo M, D’Andrea P, Nistri A, Ballerini L (2009) The patterns of spontaneous Ca2+ signals generated by ventral spinal neurons in vitro show time-dependent refinement. Eur J Neurosci 29:1543–1559CrossRefPubMedGoogle Scholar
  32. Spenger C, Braschler UF, Streit J, Luscher HR (1991) An organotypic spinal cord–dorsal root ganglion–skeletal muscle coculture of embryonic rat. I. The morphological correlates of the spinal reflex arc. Eur J Neurosci 3:1037–1053CrossRefPubMedGoogle Scholar
  33. Streit J (1993) Regular oscillations of synaptic activity in spinal networks in vitro. J Neurophysiol 70:871–878PubMedGoogle Scholar
  34. Streit J, Spenger C, Luscher HR (1991) An organotypic spinal cord–dorsal root ganglion–skeletal muscle coculture of embryonic rat. II. Functional evidence for the formation of spinal reflex arcs in vitro. Eur J Neurosci 3:1054–1068CrossRefPubMedGoogle Scholar
  35. Taccola G, Margaryan G, Mladinic M, Nistri A (2008) Kainate and metabolic perturbation mimicking spinal injury differentially contribute to early damage of locomotor networks in the in vitro neonatal rat spinal cord. Neuroscience 155:538–555CrossRefPubMedGoogle Scholar
  36. Tölle TR, Berthele A, Zieglgansberger W, Seeburg PH, Wisden W (1993) The differential expression of 16 NMDA and non-NMDA receptor subunits in the rat spinal cord and in periaqueductal gray. J Neurosci 13:5009–5028PubMedGoogle Scholar
  37. Traynelis SF, Wollmuth LP, McBain CJ, Menniti FS, Vance KM, Ogden KK, Hansen KB, Yuan H, Myers SJ, Dingledine R, Sibley D (2010) Glutamate receptor ion channels: structure, regulation, and function. Pharmacol Rev 62:405–496CrossRefPubMedGoogle Scholar
  38. Virag L, Szabo C (2002) The therapeutic potential of poly(ADP-ribose) polymerase inhibitors. Pharmacol Rev 54:375–429CrossRefPubMedGoogle Scholar
  39. Yu SW, Andrabi SA, Wang H, Kim NS, Poirier GG, Dawson TM, Dawson VL (2006) Apoptosis-inducing factor mediates poly(ADP-ribose) (PAR) polymer-induced cell death. Proc Natl Acad Sci USA 103:18314–18319CrossRefPubMedGoogle Scholar
  40. Zimmer J, Kristensen BW, Jakobsen B, Noraberg J (2000) Excitatory amino acid neurotoxicity and modulation of glutamate receptor expression in organotypic brain slice cultures. Amino Acids 19:7–21CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Neurobiology SectorInternational School for Advanced Studies (SISSA)TriesteItaly
  2. 2.Spinal Person Injury Neurorehabilitation Applied Laboratory (SPINAL)Istituto di Medicina Fisica e RiabilitazioneUdineItaly

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