Neurotoxic effects of aluminium on embryonic chick brain cultures
Abstract
Toxic damage of brain cells by aluminium (A1) is discussed as a possible factor in the development of neurodegenerative disorders in humans. To investigate neurotoxic effects of A1, serum-free cultures of mechanically dissociated embryonic chick (stage 28–29) forebrain, brain stem and optic tectum, and for comparison meningeal cells, were treated with A1 (0–1000 μM) for 7 days. Effects of A1 on cell viability (lysosomal and mitochondrial activity) and differentiation (synthesis of cell-specific proteins) were found to the brain area specific with the highest sensitivity observed in optic tectum. No inhibiting effects on cell viability could be observed in cultures of forebrain and meninges in the concentration range tested. In all three brain tissue cultures, threshold levels for the reduction of cell differentiation parameters were found at lower concentrations [concentration resulting in a 50% decrease (IC50)>180 μM] than for the inhibitionof cell viability (IC50>280 μM) indicating a specific toxic potential of A1 for cytoskeletal alterations. The culture levels of nerve cellspecific markers microtubule-associated protein type 2 (the most sensitive parameter) and the 68-kDa neurofilament were inhibited at lower concentrations (IC50 180–630 μM) than the astrocyte-specific glial fibrillary acidic protein (IC50 700–∼1000 μM), demonstrating a particularly high sensitivity of neurons in comparison to astrocytes. Based on these differences in A1 sensitivity observed for different cell markers in the various brain tissue cultures, the in vitro system used in the present study proved to be a suitable model to assess brain area and cell type-specific neurotoxic effects of A1.
Key words
Aluminium In vitro Neurotoxicity Brain Alzheimer's diseasePreview
Unable to display preview. Download preview PDF.
References
- 1.Alfrey AC (1983) Aluminum. Adv Clin Chem 23: 69–91Google Scholar
- 2.Atterwill CK, Davies WJ, Kyriakides MA (1990) an investigation of aluminium neurotoxicity using some in vitro systems. Alternat Lab Animals 18: 181–190Google Scholar
- 3.Barker FM (1992) Canthaxanthin and the eye, a critical ocular toxicological review. 1992 European meeting of the Toxicology Forum, Copenhagen (Denmark), June 1–5. CASET Ass Ltd, Fairfax (USA) pp 478–500Google Scholar
- 4.Bizzi A, Gambetti P (1986) Phosphorylation of neurofilaments is altered in aluminium intoxication. Acta Neuropathol (Berl) 71: 154–158Google Scholar
- 5.Bruinmk A (1992) Serum-free monolayer cultures of embryonic chick brain and retina: immunoassays of developmental markers, mathematical data analysis and establishment of optimal culture conditions. In: Zbinden G (ed) The brain in bits and pieces, M. T. C. Verlag,Zollikon, pp 23–50Google Scholar
- 6.Bruinink A, Birchler F (1993) Effects of cisplatin and ORG. 2766 in chick embryonic cell cultures. Arch Toxicol 67: 325–329Google Scholar
- 7.Bruinink A, Reiser P (1991) Ontogeny of MAP2 and GEAP antigens in primary cultures of embryonic chick brain. Effect of substratum, oxygen tension, serum and ARA-C. Int J Dev Neurosci 9: 269–279Google Scholar
- 8.Bruinink A, Zimmermann G, Riesen F (1991) Neurotoxic effects of chloroquine in vitro. Arch Toxicol 65: 480–484Google Scholar
- 9.Bruinink A, Reiser P, Müller M, Gähwiler BH, Zbinden G (1992) Neurotoxic effects of bismuth in vitro. Toxicol in Vitro 6: 285–293Google Scholar
- 10.Candy JM, Oakley AE, Mountfort SA, Taylor GA, Morris CM, Bishop HE, Edwardson JA (1992) The imaging and quantification of aluminium in the human brain using dynamic secondary ion mass spectrometry (SIMS). Biol Cell 74: 109–118Google Scholar
- 11.Cole GM, Wu K, Timiras PS (1985) A culture model for agerelated human neurofibrillary pathology. Int J Dev Neurosci 3: 23–32Google Scholar
- 12.Cork LC, Sternberger NH, Sternberger LA, Casanova MF, Struble RG, Price DL (1986) Phosphorylated neurofilament antigens in neurofibrillary tangles in Alzheimer's disease. J. Neuropathol Exp Neurol 45: 56–64Google Scholar
- 13.Crapper DR, Krishnan SS, Dalton AJ (1973) Brain aluminum distribution in Alzheimer's disease and experimental neurofibrillary degeneration. Science180: 511–513Google Scholar
- 14.Crapper DR, Krishnan SS, Quittkat S (1976) Aluminium, neurofibrillary degeneration and Alzheimer's disease. Brain 99: 67–80Google Scholar
- 15.Crapper McLachlan DR, Kruck TP, Lukiw WJ, Krishnan SS (1991) Would decreased aluminum ingestion reduce the incidence of Alzheimer's disease? Can Med Assoc J 145: 793–804Google Scholar
- 16.DeBoni U, Otvos A, Scott JW, Crapper DR (1976) Neurofibrillary degeneration induced by systemic aluminum. Acta Neuropathol (Berl) 35: 285–294Google Scholar
- 17.Galle P (1981) Mécanisme d'elimination rénale de deux éléments du groupe IIIA de la classification périodique: I'aluminium et l'indium. C R Acad Sci Paris 292: 91–96Google Scholar
- 18.Galle P, Berry J-P, Duckett S (1980) Electron microprobe ultrastructural localization of aluminum in rat brain. Acta Neuropathol (Berl) 49: 245–247Google Scholar
- 19.Galle P, Giudicelli CP, Nebout T, Baglin A, Fries D (1987) Ultrastructural localization of aluminum in hepatocytes of hemodialyzed patients. Ann Pathol 7: 163–170Google Scholar
- 20.Gilbert MR, Harding BL, Hoffman PN, Griffin JW, Price DL, Troncoso JC (1992) Aluminum-induced neurofilamentous changes in cultured rat dorsal root ganlia explants. J Neurosci 12: 1763–1771Google Scholar
- 21.Good PF, Perl DP, Bierer LM, Schmeidler J (1992) Selective accumulation of aluminum and iron in the neurofibrillary tangles of Alzheimer's disease: a laser microprobe (LAMMA) study. Ann Neurol 31: 286–292Google Scholar
- 22.Goodison KL, Parhad IM, White CL, Sima AA, Clark AW (1993) Neuronal and glial gene expression in neocortex of Down's syndrome and Alzheimer's disease. J Neuropathol Exp Neurol 52: 192–198Google Scholar
- 23.Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88: 49–92Google Scholar
- 24.Hewitt CC, Herman MM, Lopes MBS, Savory J, Wills MR (1991) Aluminium maltol-induced neurocytoskeletal changes in fetal rabbit midbrain in matrix culture. Neuropathol Appl Neurobiol 17: 47–60Google Scholar
- 25.Jellinger K, Braak H, Braak E, Fischer P (1991) Alzheimer lesions in the entorhinal region and isocortex in Parkinson's and Alzheimer's diseases. Ann NY Acad Sci 640: 203–209Google Scholar
- 26.Johnson GVW, Jope RS (1988) Phosphorylation of rat brain cytoskeletal proteins is increased after orally administered aluminum. Brain Res 456: 95–103Google Scholar
- 27.Johnson GVW, Watson AL, Lartius R, Uemura E, Jope RS (1992) Dietary aluminum selectively decreases MAP-2 in brains of developing and adult rats. Neurotoxicology 13: 463–474Google Scholar
- 28.Katsetos CD, Savory J, Herman MM, Carpenter RM, Frankfurter A, Hewitt CD, Wills MR (1990) Neuronal cytoskeletal lesions induced in the CNS by intraventricular and intravenous aluminium maltol in rabbits. Neuropathol Appl Neurobiol 16: 511–528Google Scholar
- 29.Kisby GE, Acosta D (1987) Cytotoxic effects of aluminum in hippocampal, cerebellar, and astrocyte cell cultures. In Vitro Toxicol 1: 85–102Google Scholar
- 30.Klatzo I, Wisniewski H, Streicher E (1965) Experimental production of neurofibrillary degeneration. J Neuropathol Expl Neurol 24: 187–199Google Scholar
- 31.Knutti R (1984) Matrixmodifikation zur direkten Bestimmung von Elementen im Ultraspurenbereich in Blut und Urin mittels Graphitrohr-Atomabsorptionsspektrometrie. In: Welz B (ed) Fortschritte in der atomspektrometrischen Spurenanalytik, vol 1. Verlag Chemie, Weinheim, pp 327–336Google Scholar
- 32.Kosik KW, Bradley WG, Goof PF, Rasool RG, Selkoe DJ (1983) Cholinergic function in lumbar aluminum myelopathy. J Neuropathol Exp Neurol 42: 365–375Google Scholar
- 33.Kosik KS, Duffy LK, Dowling MM, Abraham C, McCluskey A, Selkoe DJ (1984) Microtubule-associated protein 2: monoclonal antibodies demonstrate the selective incorporation of certain epitopes into Alzheimer neurofibrillary tangles. Proc Natl Acad Sci USA 81: 7941–7945Google Scholar
- 34.Kowall NW, Pendlebury WW, Kessler JB, Perl DP, Beal MF (1989) Alumium-induced neurofibrillary degeneration affects a subset of neurons in rabbit cerebral cortex, basal forebrain and upper brainstem. Neuroscience 29: 329–337Google Scholar
- 35.Landsberg JP, McDonald B, Watt F (1992) Absence of aluminium in neuritic plaque cores in Alzheimer's disease. Nature 360: 65–68Google Scholar
- 36.Langui D, Anderson BH, Ulrich J (1988) Effects of aluminium chloride on cultured cells from rat brain hemispheres. Brain Res 438: 67–76Google Scholar
- 37.Matus A (1988) Microtubule-associated proteins: their potential role in determining neuronal morphology. Annu Rev Neurosci 11: 29–44Google Scholar
- 38.Matus A, Bernhardt R, Hugh-Jones T (1981) High molecular weight microtubule-associated proteins are preferentially associated with dendritic microtubules in brain. Proc Natl Acad Sci USA 78: 3010–3014Google Scholar
- 39.Munoz-Garcia D, Pendlebury WW, Kessler JB, Perl DP (1986) An immunocytochemical comparison of cytoskeletal proteins in aluminum-induced and Alzheimer-type neurofibrillary tangles. Acta Neuropathol (Berl) 70: 243–248Google Scholar
- 40.Nixon RA, Clarke JF, Logvinenko KB, Tan MKH, Hoult M, Grynspan F (1990) Aluminium inhibits calpain-mediated proteolysis and induces human neurofilament proteins to form protease-resistant high molecular weight complexes. J Neurochem 55: 1950–1959Google Scholar
- 41.Norenberg MD, Norenberg LOB, Cowman GA, McCarthy M, Neary JT (1989) The effects of aluminum on astrocytes in primary culture. J Neuropathol Exp Neurol 48: 374Google Scholar
- 42.O'Callaghan JP, Jensen KF (1992) Enhanced expression of glial fibrillary acidic protein and the cupric silver degeneration reaction can be used as sensitive and early indicators of neurotoxicity. Neurotoxicology 13: 113–122Google Scholar
- 43.Perry G, Rizzuto N, Autilio-Gambetti L, Gambetti P (1985) Paired helical filaments from Alzheimer disease patients contain cytoskeletal components. Proc Natl Acad Sci USA 82: 3916–3920Google Scholar
- 44.Pierce (1990) Immuno Technology, catalog & handbook. Pierce Chemical Company, The NetherlandsGoogle Scholar
- 45.Somerville MJ, Percy ME, Bergeron C, Yoong LK, Grima EA, McLachlan DR (1991) Localization and quantitation of 68 kDa neurofilament and superoxide dismutase-1 mRNA in Alzheimer brains. Brain Res [Mol Brain Res] 9: 1–8Google Scholar
- 46.Strong MJ, Garruto RM (1991) Neuron-specific thresholds of aluminum toxicity invitro: a comparative analysis of dissociated fetal rabbit hippocampal and motor neuron-enriched cultures. Lab Invest 65: 243–249Google Scholar
- 47.Takeda M, Tatebayashi Y, Tanimukai S, Nakamura Y, Tanaka T, Nishimura T (1991) Immunohistochemical study of microtubule-associated protein-2 and ubiquitin in chronically aluminum-intoxicated rabbit brain. Acta Neuropathol 82: 346–352Google Scholar
- 48.Troncoso JC, Sternberger NH, Sternberger LA, Hoffman PN, Price DL (1986) Immunocytochemical studies of neurofilament antigens in the neurofibrillary pathology induced by aluminum. Brain Res 364: 295–300Google Scholar
- 49.Walz W (1989) Role of glial cells in the regulation of the brain ion microenvironment. Prog Neurobiol 33: 309–333Google Scholar
- 50.Wisniewski HM (1991) Neuropathology and biochemistry of Alzheimer's disease and aluminium encephalopathy. In: Detchant Lord Walton (ed) Alzheimer's disease and the environment. Royal Society of Medicine Services,London, pp 35–38Google Scholar
- 51.Zubenko GS,Hanin I (1989) Cholinergic and noradrenergic toxicity of intraventricular aluminum chloride in the rat hipocampus. Brain Res 498: 381–384Google Scholar