Experimental Brain Research

, Volume 153, Issue 3, pp 334–342 | Cite as

A peptide derived from acetylcholinesterase induces neuronal cell death: characterisation of possible mechanisms

Research Article

Abstract

Acetylcholinesterase (AChE) exhibits functions unrelated to the catalysis of acetylcholine (ACh) in particular during development. Although the underlying mechanism(s) is presently unknown, a candidate peptide fragment (AChE-peptide) has recently been identified, and been shown to induce a continuum of apoptotic and necrotic neuronal cell death in rat hippocampal organotypic cultures. The aim of this study was to trace the cell death pathway initiated by AChE-peptide. Using specific antagonists, it was possible to track a series of cellular events following application of 1 nM AChE-peptide: NMDA receptor activation, opening of the L-type voltage gated calcium channel, activation of calcium/calmodulin kinase II, generation of reactive oxygen species and caspase activation. Pharmacological interception at any stage of this cascade blocked the effect of 1 nM AChE-peptide on neurite retraction. Lactate dehydrogenase (LDH) release, a marker for cell lysis, was unaffected by 1 nM AChE-peptide. In contrast, cell death induced by 1 mM AChE-peptide, monitored as neurite retraction and increased LDH efflux, was not offset by any drug treatment. These data suggest that nanomolar concentrations of AChE-peptide exhibit pathophysiological activity via an apoptotic pathway that could play an important role in neuronal development and neurodegeneration.

Keywords

Acetylcholinesterase Cell death Hippocampus AChE-peptide 

Notes

Acknowledgements

We thank Synaptica Ltd. (Oxford, UK) for support, Dr Steven Butcher for helpful comments, and Kevin Pryor for technical assistance.

References

  1. Amoroso S, D’Alessio A, Sirabella R, Di Renzo G, Annunziato L (2002) Calcium-independent caspase-3 but not calcium-dependent caspase-2 activation induced by oxidative stress leads to SH-SY5Y human neuroblastoma cell apoptosis. J Neurosci Res 68:454–462CrossRefPubMedGoogle Scholar
  2. Anegawa NJ, Guttman RP, Grant ER, Lindstrom J, Lynch DR (2000) N-Methyl-d-aspartate receptor mediated toxicity in nonneuronal cell lines: characterisation using fluorescent measures of cell viability and reactive oxygen species. Brain Res Mol. Brain Res 77:163–175CrossRefGoogle Scholar
  3. Appleyard ME (1994) Non-cholinergic functions of acetylcholinesterase. Biochem Soc Trans 22:749–755PubMedGoogle Scholar
  4. Boutillier AL, Kienlen-Campard P, Loeffler JP (1999) Depolarisation regulates cyclin D1 degradation and neuronal apoptosis: a hypothesis about the role of the ubiquitin/proteasome signalling pathway. Eur J Neurosci 11: 441–448CrossRefPubMedGoogle Scholar
  5. Bucclantini M, Giannoni E, Fabrizio C, Baroni F, Formigli L, Zurdo J, Taddei N, Ramponi G, Dobson CM, Stefani M (2002) Inherent toxicity of aggregates implies a common mechanism for protein misfolding diseases. Nature 416:507–511CrossRefPubMedGoogle Scholar
  6. Catterall WA (2000) Structure and regulation of voltage-gated calcium channels. Annu Rev Cell Dev Biol 16:521–555PubMedGoogle Scholar
  7. Chan J, Quik M (1993) A role for the nicotinic α-bungarotoxin receptor in neurite outgrowth in PC12 cells. Neuroscience 56:441–451CrossRefPubMedGoogle Scholar
  8. Clague JR, Langer GA (1994) The pathogenesis of free radical-induced calcium leak in cultured rat cardiomyocytes. J Mol Cell Cardiol 26:11–21CrossRefPubMedGoogle Scholar
  9. Coyle JT, Puttfarcken P (1993) Oxidative stress, glutamate, and neurodegenerative disorders. Science 262:689–694PubMedGoogle Scholar
  10. Davies J, Watkins JC (1982) Actions of d and l forms of 2-amino-5-phosphovalerate and 2-amino-4-phosphobutyrate in the cat spinal cord. Brain Res 235:378–386PubMedGoogle Scholar
  11. Day T, Greenfield SA (2002) A non-cholinergic, trophic action of acetylcholinesterase on hippocampal neurones in vitro: molecular mechanisms. Neuroscience 111:649–656CrossRefPubMedGoogle Scholar
  12. Day T, Greenfield SA (2003) Bioactivity of a peptide derived from acetylcholinesterase in hippocampal organotypic cultures. Exp Brain Res (in press)Google Scholar
  13. De Garcia DJ, Kumar R, Owen CR, Krause GS, White BC (2002) Molecular pathways of protein synthesis inhibition during brain reperfusion: implications for neuronal survival or death. J Cereb. Blood Flow Metab 22:127–141PubMedGoogle Scholar
  14. Dickie BGM, Holmes C, Greenfield SA (1996) Neurotoxic and neurotrophic effects of chronic NMDA exposure upon mesencephalic dopaminergic neurons in organotypic culture. Neuroscience 72:731–741CrossRefPubMedGoogle Scholar
  15. Dykens JA (1994) Isolated cerebral and cerebellar mitichondria produce free radicals when exposed to elevated calcium and sodium: implications for neruodegeneration. J Neurochem 63:584–591PubMedGoogle Scholar
  16. Eimerl S, Schramm M (1994) The quality of calcium that appears to induce neuronal death. J Neurochem 62:1223–1226PubMedGoogle Scholar
  17. Ferri KF, Kroemer G (2001) Organelle-specific initiation of cell death pathways. Nat Cell Biol 3:E255–E263Google Scholar
  18. Gähwiler BH (1981) Organotypic monolayer cultures of nervous tissue. J Neurosci Methods 4:329–342PubMedGoogle Scholar
  19. Gähwiler B.H (1984) Development of the hippocampus in vitro: cell types, synapses and receptors. Neuroscience 11:751–760CrossRefPubMedGoogle Scholar
  20. Giovanni A, Wirtz-Brugger F, Keramaris E, Slack R, Park DS (1999) Involvement of cell cycle elements, cyclin-dependent kinases, pRb, and E2F-DP, in B-amyloid-induced neuronal death. J Biol Chem 274:19011–19016CrossRefPubMedGoogle Scholar
  21. Greenfield SA (1996) Non-classical actions of acetylcholinesterase: role in cellular differentiation, tumorigenesis and Alzheimer’s disease: a critique. Neurochem Int 28:485–490CrossRefPubMedGoogle Scholar
  22. Greenfield SA, Vaux D (2002) Parkinson’s disease, Alzheimer’s disease and motor neuron disease: identifying a common mechanism. Neuroscience 113:485–492CrossRefPubMedGoogle Scholar
  23. Grundman M (2000) Vitamin E and Alzheimer’s disease: the basis for additional clinical trials. Am. J Clin Nutr 71 [Suppl]:630s–636sGoogle Scholar
  24. Guo Q, Sebastain L, Sopher BL, Miller MW, Ware CB, Martin GM, Mattson MP (1999) Increased vulnerability of hippocampal neurons from presenilin-1 mutant knock-in mice to amyloid β-peptide toxicity: Central roles of superoxide production and caspase activation. J Neurochem 72:1019–1029CrossRefPubMedGoogle Scholar
  25. Han BH, Xu D, Choi J, Han Y, Xanthoudakis S, Roy S, Tam J, Vaillancourt J, Colucci J, Siman R, Giroux A, Robertson GS, Zamboni R, Nicholson DW, Holtzman DM (2002) Selective, reversible caspase-3 inhibitor is neuroprotective and reveals distinct pathways of cell death after neonatal hypoxic-ischemic brain injury. J Biol Chem 277:30128–30136CrossRefPubMedGoogle Scholar
  26. Harada J, Sugimoto M (1999) Activation of caspase-3 in β-amyloid-induced apoptosis of cultured rat cortical neurons. Brain Res 842:311–323CrossRefPubMedGoogle Scholar
  27. Heng J.E, Zurakowski D, Vorwerk C.K, Grosskreutz C.L, Dreyer E.B (1999) Cation channel control of neurite morphology. Brain Res Dev Brain Res 113:67–73CrossRefPubMedGoogle Scholar
  28. Holmes C, Jones SA, Budd TC, Greenfield SA (1996) A non-cholinergic, trophic action of recombinant acetylcholinesterase on mid-brain dopaminergic neruons. J Neurosci Res 49:1–12Google Scholar
  29. Huang H, Ou H, Hseih S (2000) Antioxidiants prevent amyloid peptide-induced apoptosis and alteration of calcium homeostasis in cultured cortical neurons. Life Sci 66:1879–1892CrossRefPubMedGoogle Scholar
  30. Jacobson MD (1996) Reactive oxygen species and programmed cell death. Trends Biochem Sci 21:83–86CrossRefPubMedGoogle Scholar
  31. Jellinger KA, Stadelmann C (2001) Problems of cell death in neurodegeneration and Alzheimer’s disease. J Alzheimers Dis 3:31–40PubMedGoogle Scholar
  32. Joseph JA, Strain JG, Jimenez ND, Fisher D (1997) Oxidant injury on PC12 cells—A possible model of calcium “dysregulation” in ageing: I. Selectivity of protection against oxidative stress. J Neurochem 69:1252–1258PubMedGoogle Scholar
  33. Kim PK, Kwon YG, Chung HT, Kim YM (2002) Regulation of caspases by nitric oxide. Ann NY Acad Sci 962:42–52PubMedGoogle Scholar
  34. Le WD, Colom LV, Xie WJ, Smith G, Alexianu M, Appel SH (1995) Cell death induced by β amyloid 1–40 in MES 23.5 hybrid clone: the role of nitric oxide and NMDA-gated channel activation leading to apoptosis. Brain Res 686:49–60CrossRefPubMedGoogle Scholar
  35. Lei SZ, Pan Z, Aggarwal SK, Chen HV, Hartman J, Sucher NJ, Lipton SA (1992) Effect of nitric oxide production on the redox modulatory site of the NMDA receptor channel complex. Neuron 8:1087–1099PubMedGoogle Scholar
  36. Lukas RJ, Lucero L, Buisson B, Galzi JL, Puchacz E, Fryer JD, Changeaux JP, Bertrand D (2001) Neurotoxicity of channel mutations in heterologously expressed α7-nicotinic acetylcholine receptor. Eur J Neurosci 13:1849–1860CrossRefPubMedGoogle Scholar
  37. Marshall K, Reitter R.J, Poeggeler B, Aruoma OI, Halliwell B (1996) Evaluation of the antioxidant activity of melatonin in vitro. Free Radic Biol Med 21: 307–315CrossRefPubMedGoogle Scholar
  38. Mironov SL, Richter DW (2000) Hypoxic modulation of L-type calcium channels in inspiratory brainstem neurones: intracellular signalling pathways and metabotropic glutamate receptors. Brain Res 869:166–177PubMedGoogle Scholar
  39. Monji A, Utsumi H, Ueda T, Imoto T, Yoshida I, Hashioka S, Tashiro K, Tashiro N (2001) The relationship between the aggregational state of the β-amyloid peptides and free radical generation by the peptides. J Neurochem 77:1425–1432CrossRefPubMedGoogle Scholar
  40. Montal M (1998) Mitochondria, glutamate neurotxicity and the death cascade. Biochim Biophys Acta 1366:113–126CrossRefPubMedGoogle Scholar
  41. Moore JD, Rothwell NJ, Gibson RM (2002) Involvement of caspases and calpains in cerebrocortical neuronal cell death is stimulus-dependent. Br J Pharmacol 135:1069–1077PubMedGoogle Scholar
  42. Moya E, Blagbrough IS (1996) Efficient syntheses of polyamine and polyamine amide voltage-sensitive calcium channel blockers: FTX-3.3 and sFTX-3.3. J Pharm Pharmacol 48:179–182PubMedGoogle Scholar
  43. Nagy ZS, Esiri MM, Smith AD (1998) The cell division cycle and the pathophysiology of Alzheimer’s disease. Neuroscience 87:731–739Google Scholar
  44. Ohkuma S, Katsura M, Higo K, Hara A, Tarumi C, Ohgi T (2001) Peroxynitrite affects calcium influx through voltage-dependent calcium channels. J Neurochem 76:341–350CrossRefPubMedGoogle Scholar
  45. Okabe E, Odajima C, Taga R, Kukreja R.C, Hess ML, Ito H (1988) The effect of oxygen free radicals on calcium permeability and calcium loading at steady state in cardiac sarcoplasmic reticulum. Mol Pharmacol 34:388–394PubMedGoogle Scholar
  46. Park DS, Morris EJ, Greene LA, Geller HM (1997) G1/S cell cycle blockers and inhibitors of cyclin-dependent kinases suppress camptothecin-induced neuronal apoptosis. J Neurosci 17:1256–1270PubMedGoogle Scholar
  47. Perez-Velazquez JL, Frantseva MV, Carlen PL (1997) In vitro ischemia promotes glutamate-mediated free radical generation and intracellular calcium accumulation in hippocampal pyramidal neurons. J Neurosci 17:9085–9094PubMedGoogle Scholar
  48. Pugh PC, Berg DK (1994) Neuronal acetylcholine receptors that bind α-bungarotoxin mediate neurite retraction in a calcium-dependent manner. J Neurosci 14:889–896PubMedGoogle Scholar
  49. Raina AK, Monteiro MJ, McShea A, Smith MA (1999) The role of cell cycle-mediated events in Alzheimer’s disease. Int J Exp Pathol 80:71–76CrossRefPubMedGoogle Scholar
  50. Raina AK, Zhu X, Rottkamp CA, Monteiro M, Takeda A, Smith MA (2000) Cyclin’ toward dementia: cell cycle abnormalities and abortive oncogenesis in Alzheimer disease. J Neurosci Res 61:128–133CrossRefPubMedGoogle Scholar
  51. Ramassamy C, Averill D, Beffert U, Theroux L, Lussier-Cacon S, Cohn JS, Christen Y, Schoofs A, Davignon J, Poirier J (2000) Oxidative insults are associated with apolipoprotein E genotype in Alzheimer’s disease brain. Neurobiol Dis 7:23–37CrossRefPubMedGoogle Scholar
  52. Reiter R, Tang L, Garcia JJ, Muñoz-Hoyos (1997) Pharmacological actions of melatonin in oxygen radical pathophysiology. Life Sci 60:2255–2271PubMedGoogle Scholar
  53. Reiter R, Tan D, Cabrera J, D’Arpa D, Sainz RM, Mayo JC, Ramos S (1999) The oxidant/antioxidant network: role of melatonin. Biol Signals Recept 8:56–63CrossRefPubMedGoogle Scholar
  54. Sakai K, Suzuki K, Tanaka S, Koike T (1999) Up-regulation of cylcin D1 occurs in apoptosis of immature but not mature cerebellar granule neurons in cultures. J Neurosci Res 58:396–406PubMedGoogle Scholar
  55. Séguéla P, Wadiche J, Dineley-Miller K, Dani JA, Patrick JW (1993) Molecular cloning, functional properties and distribution of rat brain α7: a nicotinic cation channel highly permeable to calcium. J Neurosci 13:596–604PubMedGoogle Scholar
  56. Soderling TR, Fukunaga K, Rich DP, Fong YL, Smith K, Colbran RJ (1990) Regulation of brain Ca2+/calmodulin-dependent protein kinase II. Adv Second Messenger Phosphoprotein Res 24:206–211PubMedGoogle Scholar
  57. Sombati S, Coulter DA, DeLorenzo RJ (1991) Neurotoxic activation of glutamate receptors induces an extended neuronal depolarisation in cultured hippocampal neurons. Brain Res 566:316–319CrossRefPubMedGoogle Scholar
  58. Spedding M, Paoletti R (1992) III. Classification of calcium channels and the sites of action of drugs modifying channel function. Pharmacol Rev 44:363–376PubMedGoogle Scholar
  59. Ueda K, Shinohara S, Yagami T, Asakura K, Kawasaki K (1997) Amyloid β protein potentiates calcium influx through L-type voltage sensitive calcium channels: a possible involvement of free radicals. J Neurochem 68:265–271PubMedGoogle Scholar
  60. Webb CP, Nedergaard S, Giles K, Greenfield SA (1996) Involvement of the NMDA receptor in a non-cholinergic action of acetylcholinesterase in guinea-pig substantia nigra pars compacta neurons. Eur J Neurosci 8:837–841PubMedGoogle Scholar
  61. Zhang C, Shen W, Zhang G (2002) N-methyl-d-aspartate receptor and L-type voltage-gated calcium channel antagonists suppress the release of cytochrome c and the expression of procaspase-3 in rat hippocampus after global brain ischemia. Neurosci Lett 328:265–268CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Department of PharmacologyUniversity of OxfordOxfordUK

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