Neurotoxicity Research

, Volume 9, Issue 4, pp 297–304 | Cite as

Cerebellar Granule Cell Death Induced by Aluminum

  • Jelena Tuneva
  • Sridar Chittur
  • Alexander A. Boldyrev
  • Irina Birman
  • David O. Carpenter


Using flow cytometry of acutely isolated cerebellar granule cell neurons, we have determined the effects of Al3+ on viability, membrane potential, intracellular calcium concentration and generation of reactive oxygen species (ROS). Al3+ killed granule cells in a time- and concentration-dependent fashion when monitored by use of the DNA-binding dye, propidium iodide. The threshold concentration was about 50 µM, and cell death at 100 µM was apparent after 30 min exposure and increased over time. Cell death was accompanied by cell swelling and a decrease in membrane potential, and was not dependent on external calcium concentration. While exposure to Al3+ was accompanied by an increase in ROS and an elevation of intracellular calcium concentration, calcium chelators and ROS scavengers did not reduce cell death. The action of Al3+ was not accompanied by activation of caspase-3 or an increase in annexin-V binding, both indicators of apoptosis. In the presence of intracellular O,O’-bis(2-aminophenyl)ethyleneglycol-N,N,N’,N’-tet-raacetic acid (BAPTA) and absence of extracellular calcium there was still a fluo-3 signal, which likely reflects an accumulation of intracellular Al3+. These observations suggest that the cell death is subsequent to intracellular accumulation of Al3+ and subsequent perturbation of cellular metabolism.


Aluminum Oxidative stress Reactive oxygen species Intracellular calcium N-acetylcysteine Cerebellar granule cells Flow cytometry 


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  1. Alfrey AC (1993) Aluminum and renal disease, In:Moving Points in Nephrology (Bourke E, NP Mallick and VE Pollack, Eds.) (Karger:Basel), pp 110–124.Google Scholar
  2. Alshuaib WB, SP Cherian, MY Hasan and MA Fahim (2003) Drug effects on calcium homeostasis in mouse CA1 hippocampal neurons.Int. J. Neurosci. 113, 1317–1332.PubMedCrossRefGoogle Scholar
  3. Altmann P, J Cunningham, U Dhanesha, M Ballard, J Thompson and F Marsh (1999) Disturbance of cerebral function in people exposed to drinking water contaminated with aluminum sulphate: retrospective study of the Camelford water incident.BMJ 319, 807–811.PubMedGoogle Scholar
  4. Bertholf RL, JR Nicholson, MR Wills and J Savoy (1987) Measurement of lipid peroxidation products in rabbit brain and organs (response to aluminum exposure).Ann. Clin. Lab. Sci. 17, 418–423.PubMedGoogle Scholar
  5. Boldyrev A, R Song, VA Dyatlov, DA Lawrence and DO Carpenter (2000) Neuronal cell death and reactive oxygen species.Cell. Molec. Neurobiol. 20, 433–450.PubMedCrossRefGoogle Scholar
  6. Boldyrev A, V Kazey, T Leinsoo, A Mashkina, O Tyulina, P Johnson, O Tuneva, S Chittur and A Boldyrev (2004) Rodent lymphocytes express functionally active glutamate receptors.Biochem. Biophys. Res. Commun. 324, 133–139.PubMedCrossRefGoogle Scholar
  7. Bondy SC, SF Ali and S Guo-Ross (1998) Aluminum but not iron treatment induces pro-oxidant events in the rat brain.Mol. Chem. Neuropathol. 34, 219–232.PubMedCrossRefGoogle Scholar
  8. Brenner S (2002) Aluminum neurotoxicity is reduced by dantrolene and dimethyl sulfoxide in cultured rat hippocampal neurons.Biol. Trace Element Res. 86, 85–89.CrossRefGoogle Scholar
  9. California Environmental Protection Agency (2001) Public Health Goal for ALUMINUM in Drinking Water. (accessed 14 April 2005).Google Scholar
  10. Campbell A, MA Smith, LM Sayre, SC Bondy and G Perry (2001) Mechanisms by which metals promote events connected to neurodegenerative diseases.Brain Res. Bull. 55, 125–132.PubMedCrossRefGoogle Scholar
  11. Carpenter DO, CR Stoner and D Lawrence (1997) Flow cytometry measurements of neuronal cell death triggered by PCBs.NeuroToxicology 18, 507–514.PubMedGoogle Scholar
  12. Chabannes E, S Fauconnet, S Bernardini, H Wallerand, G Adessi and H Bittard (2001) Protein kinase C signaling pathway is involved in the regulation of vascular endothelial growth factor expression in human bladder transitional carcinoma cells.Cell Signal 13, 585–591.PubMedCrossRefGoogle Scholar
  13. Charlot JF, JL Pretet, C Haughey and C Mougin (2004) Mitochondrial translocation of p53 and mitochondrial membrane potential (Delta Psi m) dissipation are early events in staurosporine-induced apoptosis of wild type and mutated p53 epithelial cells.Apoposis 9, 333–343.CrossRefGoogle Scholar
  14. El-Fattah AA, HM Al-Yousef, AM Al-Bekairi and HA Al-Sawaf (1998) Vitamin E protects the brain against oxidative injury stimulted by excessive aluminum intake.Biochim. Molec. Biol. Intl. 46, 1175–1180.Google Scholar
  15. Esparza JL, M Gomez, M Romeu, M Mulero, DJ Sanchez, J Mollol and JL Domingo (2003) Aluminum-induced pro-oxidant effects in rats: protective role of exogenous melatonin.J. Pineal Res. 35, 32–39.PubMedCrossRefGoogle Scholar
  16. Farina M, LN Rotta, FA Soares, F Jardim, R Jacques, DO Souza and JB Rocha (2005) Hematological changes in rats chronically exposed to oral aluminum.Toxicology 209, 29–37.PubMedCrossRefGoogle Scholar
  17. Flaten TP (1990) Geographical associations between aluminum in drinking water and death rates with dementia (including Alzheimer’s disease), Parkinson’s and amyotrophic lateral sclerosis in Norway.Environ. Geochem. Health 12, 152–167.CrossRefGoogle Scholar
  18. Flaten TP (2001) Aluminum as a risk factor in Alzheimer’s disease, with emphasis on drinking water.Brain Res. Bull. 55, 187–196.PubMedCrossRefGoogle Scholar
  19. Forbes WF and GB Hill (1998) Is exposure to aluminum a risk factor for the development of Alzheimer Disease?---Yes.Arch. Neurol. 55, 740–741.PubMedCrossRefGoogle Scholar
  20. Fu HJ, QSK Hu, ZN Lin, TL Ren, H Song, CK Cai and SZ Dong (2003) Aluminum-induced apoptosis in cultured cortical neurons and its effect on SAPK/JNK signal transduction pathway.Brain Res. 980, 11–23.PubMedCrossRefGoogle Scholar
  21. Gauthier E, I Fortier, F Courchesne, P Pepin, J Mortimer and D Gauvreau (2000) Aluminum forms in drinking water and risk of Alzheimer’s disease.Environ. Res. A 84, 234–246.CrossRefGoogle Scholar
  22. Gomez M, JL Esparza, MR Nogues, M Ciralt, M Cabre and JL Domingo (2005) Pro-oxidant activity of aluminum in the rat hippocampus: gene expression of antioxidant enzymes after melatonin administration.Free Radic. Biol. Med. 38, 104–111.PubMedCrossRefGoogle Scholar
  23. Griffioen KJ, O Ghribi, N Fox, J Savory and DA DeWitt (2004) Aluminum maltolate-induced toxicity in NT2 cells occurs through apoptosis and includes cytochrome c release.NeuroToxicology 25, 859–867.PubMedCrossRefGoogle Scholar
  24. Guo GW and YX Liang (2001) Aluminum-induced apoptosis in cultured astrocytes and its effect on calcium homeostasis.Brain Res. 888, 221–226.PubMedCrossRefGoogle Scholar
  25. Gupta VB, S Anitha, ML Hegde, L Zecca, RM Barruto, R Ravid, SK Shankar, R Stein, P Shanmugavelu and KSJ Rao (2005) Aluminium in Alzheimer’s disease: are we still at a crossroad?Cell. Mol. Life Sci. 62, 143–158.PubMedCrossRefGoogle Scholar
  26. Haugland RP (2002)Handbook of Fluorescent Probes and Research Products, 9th Edition (Gregory J and MTZ Spence, Eds.) (Molecular Probes, Inc.:Oregon), 965 pp.Google Scholar
  27. Johnson GV, KW Cogdill and RS Jope (1990) Oral aluminum altersin vitro protein phosphorylation and kinase activities in rat brain.Neurobiol. Aging 11, 209–216.PubMedCrossRefGoogle Scholar
  28. Johnson VJ, M Tsunoda, TF Murray and RP Sharma (2005) Decreased membrane fluidity and hyperpolarization in aluminum-treated PC-12 cells correlates with increased production of cellular oxidants.Environ. Toxicol. Pharmacol. 19, 221–230.CrossRefGoogle Scholar
  29. Kaneko N, H Yasui, J Takada, K Suzuki and H Sakurai (2004) Orally administrated aluminum-maltolate complex enhances oxidative stress in the organs of mice.J. Inorg. Biochem. 98, 2022–2031.PubMedCrossRefGoogle Scholar
  30. Landsberg JP, B McDonald and F Watt (1992) Absence of aluminum in neuritic plaque cores in Alzheimer’s disease.Nature 360, 65–67.PubMedCrossRefGoogle Scholar
  31. Lankoff A, A Banasik, A Duma, E Ochniak, H Lisowska, T Kuszewski, S Gozdz and A Wojcik (2006) A comet assay study reveals that aluminum induces DNA damage and inhibits the repair of radiation-induced lesions in human peripheral blood lymphocytes.Toxicol. Lett. 161, 27–36.PubMedCrossRefGoogle Scholar
  32. Martyn CN, C Osmond, JA Edwardson, DJP Barker, EC Harris and RF Lacey (1989) Geographical relation between Alzheimer’s Disease and aluminum in drinking water.Lancet 1, 60–62.Google Scholar
  33. Moumen R, N Ait-Oukhatar, F Bureau, C Fleury, D Bougle, P Arhan, D Neuville and F Viader (2001) Aluminum increases xanthine oxidase activity and disturbs antioxidant status in the rat.J. Trace Elem. Med. Biol. 15, 89–93.PubMedCrossRefGoogle Scholar
  34. Munoz DG (1998) Is exposure to aluminum a risk factor for the development of Alzheimer disease?---No.Arch. Neurol. 55, 737–739.PubMedCrossRefGoogle Scholar
  35. Nehru B and P Anand (2005) Oxidative damage following chronic aluminum exposure in adult and pup rat brains.J. Trace Elem. Med. Biol. 19, 203–208.PubMedGoogle Scholar
  36. Oteiza PI, CG Frage and CL Keen (1993) Aluminum has both oxidant and antioxidant effects in mouse brain membranes.Arch. Biochem. Biophys. 300, 517–521.PubMedCrossRefGoogle Scholar
  37. Oyama Y, DO Carpenter, L Chikahisa and E Okazaki (1996) Flowcytometric estimation on glutamate- and kainite-induced increases in intracellular Ca2+ of brain neurons: a technical aspect.Brain Res. 728, 121–124.PubMedCrossRefGoogle Scholar
  38. Perl DP and AR Brody (1980) Alzheimer’s disease: X-ray spectrometric evidence of aluminum accumulation in neurofibrillary tangle-bearing neurons.Science 208, 297–299.PubMedCrossRefGoogle Scholar
  39. Platt B, DO Carpenter, D Busselberg, KG Reymann and G Riedel (1995) Aluminum impairs hippocampal long-term potentiation in ratsin vitro andin vivo. Exp. Neurol. 134, 73–86.CrossRefGoogle Scholar
  40. Provan SD and RA Yokel (1992) Aluminum inhibits glutamate release from transverse rat hippocampal slices: role of G proteins, Ca channels and protein kinase C.NeuroToxicology 13, 413–420.PubMedGoogle Scholar
  41. Rabe A, MH Lee, J Shek and HM Wisniewski (1982) Learning deficit in immature rabbits with aluminum-induced neurofibrillary changes.Exp. Neurol. 76, 441–446.PubMedCrossRefGoogle Scholar
  42. Shin RW, VM Lee and JQ Trojanowski (1994) Aluminum modifies the properties of Alzheimer’s disease PHF tau proteinsin vivo andin vitro. J. Neurosci. 14, 7221–7233.Google Scholar
  43. Silva VS, JM Cordeiro, MJ Matos, CR Oliveira and PP Goncalves (2002) Aluminum accumulation and membrane fluidity alteration in synaptosomes isolated from rat brain cortex following aluminum ingestion: effect of cholesterol.Neurosci. Res. 44, 181–193.PubMedCrossRefGoogle Scholar
  44. Singh R, R Beriault, J Middaugh, R Hamel, D Chenier, VD Appanna and S Kalyuzhnyi (2005) Aluminum-tolerant Pseudomonas fluorescens: ROS toxicity and enhanced NADPH productionExtremophiles 9, 367–373.PubMedCrossRefGoogle Scholar
  45. Trump BF and IK Berezesky (1995) Calcium-mediated cell injury and cell death.FASEB J. 9, 219–228.PubMedGoogle Scholar
  46. Tsien RY (1980) New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures.Biochemistry 19, 2396–2404.CrossRefGoogle Scholar
  47. Tsubouchi R, HH Htay, K Murakami, M Haneda and M Yoshino (2001) Aluminum-induced apoptosis in PC12D cells. Biometals14, 181–185.PubMedCrossRefGoogle Scholar
  48. Tyurina YY, VA Tyurin, Q Zhao, M Djukic, PJ Quinn, BR Pitt and VE Kagan (2004) Oxidation of phosphatidylserine: a mechanism for plasma membrane phospholipid scrambling during apoptosis?Biochem. Biophys. Res. Commun. 324, 1059–1064.PubMedCrossRefGoogle Scholar
  49. Valencia A and J Moran (2004) Reactive oxygen species induce different cell death mechanisms in cultured neurons.Free Radic. Biol.Med. 36, 1112–1125.PubMedCrossRefGoogle Scholar
  50. Wang M, DY Ruan, JT Chen and YZ Xu (2002) Lack of effects of vitamin E on aluminum-induced deficit of synaptic plasticity in rat dentate gyrusin vivo. Food Chem.Toxicol. 40, 471–478.CrossRefGoogle Scholar
  51. WHO (World Health Organization) (1997)Environmental Health Criteria 194. Aluminum. Geneva: 1997.Google Scholar
  52. Yamamoto Y, Y Kobayashi, SR Devi, S Rikiishiu and H Matsumoto (2002) Aluminum toxicity is associated with mitochondrial dysfunction and the production of reactive oxygen species in plant cells.Plant Physiol. 128, 63–72.PubMedCrossRefGoogle Scholar
  53. Zimmermann B, AV Somlyo, GC Ellis-Davies, JH Kaplan and AP Somlyo (1995) Kinetics of prephosphorylation reactions and myosin light chain phosphorylation in smooth muscle. Flash photolysis studies with caged calcium and caged ATP.J. Biol. Chem. 270, 23966–23974.PubMedCrossRefGoogle Scholar

Copyright information

© FP Graham Publishing Co 2006

Authors and Affiliations

  • Jelena Tuneva
    • 1
  • Sridar Chittur
    • 2
  • Alexander A. Boldyrev
    • 1
    • 3
  • Irina Birman
    • 1
  • David O. Carpenter
    • 1
  1. 1.Institute for Health and the EnvironmentUniversity at AlbanySUNY, RensselaerUSA
  2. 2.Center for Functional GenomicsUniversity at AlbanySUNY, RensselaerUSA
  3. 3.Center for Molecular Medicine and International Biotechnological CenterM.V. Lomonosov Moscow State UniversityMoscowRussia

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