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Cerebrospinal fluid from amyotrophic lateral sclerosis has no effect on intracellular free calcium in cultured cortical neurons

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Molecular and Chemical Neuropathology

Abstract

The data from the literature regarding the presence of a neurotoxic factor in amyotrophic lateral sclerosis (ALS) plasma or cerebrospinal fluid (CSF) remain controversial. As a new approach to this question, we have studied the effect of CSF from ALS patients on the temporal dynamics of the intracellular free calcium concentration ([Ca2+]i) of murine cortical neurons in cultures using Fura-2 fluorescence videomicroscopy and single-cell imaging. CSF from seven ALS patients and controls was added at dilutions up to 20% to cortical neuronal cultures. The in vitro inhibition of CSF on [3H]kainic acid binding showed that the CSF did not contain any substances other than glutamate itself in larger amounts. At the concentrations used, the CSF did not have any effect on [Ca2+]i or on the neuronal responsiveness as defined by the ability of the cells to respond with a transient increase in [Ca2+]i to depolarization induced by KCl. The disturbance of the intracellular calcium homeostasis is one of the key mechanisms of action of excitotoxic compounds mediating delayed neuronal cell death by stimulation of glutamate receptor subtypes. In this study, CSF from ALS patients did not induce immediate rises in [Ca2+]i or disturbances of the intracellular calcium homeostasis when measured over a period of 2 h.

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References

  • Askanas V., Marangos P. J., and Engel W. K. (1981) CSF from amyotrophic lateral sclerosis patients applied to motor neurons in culture fails to alter neuron-specific enolase.Neurology 31, 1196–1197.

    PubMed  CAS  Google Scholar 

  • Blin O., Samuel D., Nieoullon A., and Serratice G. (1994) Changes in CSF amino acid concentrations during the evolution of amyotrophic lateral sclerosis.J. Neurol. Neurosurg. Psychiatry 57, 119–120.

    Article  PubMed  CAS  Google Scholar 

  • Camu W., Billiard M., and Baldy-Moulinier M. (1993) Fasting plasma and CSF amino acid levels in amyotrophic lateral sclerosis: a subtype analysis.Acta Neurol. Scand. 88, 51–55.

    Article  PubMed  CAS  Google Scholar 

  • Couratier P., Hugon J., Sindou P., Vallat J. M., and Dumas M. (1993) Cell culture evidence for neuronal degeneration in amyotrophic lateral sclerosis being linked to glutamate AMPA/kainate receptors.Lancet 341, 265–268.

    Article  PubMed  CAS  Google Scholar 

  • Couratier P., Sindou P., Esclaire F., Louvel E., and Hugon J. (1994) Neuroprotective effects of riluzole in ALS CSF toxicity.Neuroreport 5, 1012–1014.

    Article  PubMed  CAS  Google Scholar 

  • Frandsen A. and Schousboe A. (1987) Time and concentration dependency of toxicity of excitatory amino acids on cerebral neurons in primary culture.Neurochem. Int. 10, 583–591.

    Article  CAS  PubMed  Google Scholar 

  • Frandsen A. and Schousboe A. (1990) Development of excitatory amino acid induced cytotoxicity in cultured neurons.Int. J. Dev. Neurosci. 8, 209–216.

    Article  PubMed  CAS  Google Scholar 

  • Frandsen A. and Schousboe A. (1991) Dantrolene prevents glutamate cytotoxicity and Ca2+ release from intracellular stores in cultured cerebral cortical neurons.J. Neurochem. 56, 1075–1078.

    Article  PubMed  CAS  Google Scholar 

  • Frandsen A. and Schousboe A. (1993) Excitatory amino acid-mediated cyto-toxicity and calcium homeostasis in cultured neurons.J. Neurochem. 60, 1202–1211.

    Article  PubMed  CAS  Google Scholar 

  • Honoré T., Drejer J., and Nielsen M. (1986) Calcium discriminates two3H-kainate binding sites with different molecular target sizes in rat cortex.Neurosci. Lett. 65, 47–52.

    Article  PubMed  Google Scholar 

  • Horwich M. S., Engel W. K., and Chauvin P. B. (1974) Amyotrophic lateral sclerosis sera applied to cultured motor neurons.Arch. Neurol. 30, 332–333.

    PubMed  CAS  Google Scholar 

  • Lehrich J. R. and Couture J. (1978) Amyotrophic lateral sclerosis sera are not cytotoxic to neuroblastoma cells in tissue culture.Ann. Neurol. 4, 384.

    Article  PubMed  CAS  Google Scholar 

  • Liveson J., Frey H., and Bornstein M. B. (1975) The effect of serum from ALS patients on organotypic nerve and muscle, tissue cultures.Acta Neuropathol. Berl. 32, 127–131.

    Article  PubMed  CAS  Google Scholar 

  • Marti-Fàbregas J., Soler R., Esquerda J., Grau J. M., Pradas J., and Illa I. (1992) Clinical status of motoneuron disease does not correlate with serum neurotoxicity on cultured neurons.Acta Neurol. Scand. 85, 219–223.

    Article  PubMed  Google Scholar 

  • Møller S. E. (1993) Quantification of physiological amino acids by gradient ion-exchange high-performance liquid chromatography.J. Chromatogr. 613, 223–230.

    Article  PubMed  Google Scholar 

  • Papageorgiou C. T., Libitaki G. S., Kapaki E. N., Zournas C. P., and Segditsa I. T. (1993) Amino acid concentration in the cerebrospinal fluid in amyotrophic lateral sclerosis.Med. Sci. Res. 21, 561–562.

    CAS  Google Scholar 

  • Patten B. M., Harati Y., Acosta L., Jung S. S., and Felmus M. T. (1978) Free amino acid levels in amyotrophic lateral sclerosis.Ann. Neurol. 3, 305–309.

    Article  PubMed  CAS  Google Scholar 

  • Perry T. L., Krieger C., Hansen S., and Eisen A. (1990) Amyotrophic lateral sclerosis: amino acid levels in plasma and cerebrospinal fluid.Ann. Neurol. 28, 12–17.

    Article  PubMed  CAS  Google Scholar 

  • Plaitakis A. and Caroscio J. T. (1987) Abnormal glutamate metabolism in amyotrophic lateral sclerosis.Ann. Neurol. 22, 575–579.

    Article  PubMed  CAS  Google Scholar 

  • Randall R. D. and Thayer S. A. (1992) Glutamate-induced calcium transient triggers delayed calcium overload and neurotoxicity in rat hippocampal neurons.J. Neurosci. 12, 1882–1895.

    PubMed  CAS  Google Scholar 

  • Roisen F. J., Bartfeld H., Donnenfeld H., and Baxter J. (1982) Neuron specific in vitro cytotoxicity of sera from patients with amyotrophic lateral sclerosis.Muscle Nerve 5, 48–53.

    Article  PubMed  CAS  Google Scholar 

  • Rothstein J. D., Tsai G., Kuncl R. W., Clawson L., Cornblath D. R., Drachman D. B., Pestronk A., Stauch B. L., and Coyle J. T. (1990) Abnormal excitatory amino acid metabolism in amyotrophic lateral sclerosis.Ann. Neurol. 28, 18–25.

    Article  PubMed  CAS  Google Scholar 

  • Rothstein J. D., Kuncl R., Chaundhry V., Clawson L., Cornblath D. R., Coyle J. T., and Drachman D. B. (1991) Excitatory amino acids in amyotrophic lateral sclerosis: an update.Ann. Neurol. 30, 224–225.

    Article  PubMed  CAS  Google Scholar 

  • Rothstein J. D., Martin L. J., and Kuncl R. W. (1992) Decreased glutamate transport by the brain and spinal cord in amyotrophic lateral sclerosis.N. Engl. J. Med. 326, 1464–1468.

    Article  PubMed  CAS  Google Scholar 

  • Silani V., Pizzuti A., Redaelli L. M., Bassani R., Causarano I. R., Buscaglia M., Zuliani G., and Scarlato G. (1987) ALS cerbrospinal fluid enhances human foetal astroglial cell proliferation in vitro.Adv. Exp. Med. Biol. 209, 79–81.

    PubMed  CAS  Google Scholar 

  • Touzeau G. and Kato A. C. (1983) Effects of amyotrophic lateral sclerosis sera on cultured cholinergic neurons.Neurology 33, 317–322.

    PubMed  CAS  Google Scholar 

  • Touzeau G. and Kato A. C. (1986) ALS serum has no effect on three enzymatic activities in cultured human spinal cord neurons.Neurology 36, 573–576.

    PubMed  CAS  Google Scholar 

  • Tymianski M., Charlton M. P., Carlen P. L., and Tator C. H. (1993) Source specificity of early calcium neurotoxicity in cultured embryonic spinal neurons.J. Neurosci. 13, 2085–2104.

    PubMed  CAS  Google Scholar 

  • Witt M. R., Dekermendjian K., Frandsen A., Schousboe A., and Nielsen M. (1994) Complex correlation between excitatory amino acid-induced increase in the intracellular Ca2+ concentration and subsequent loss of neuronal function in individual neocortical neurons in culture.Proc. Natl. Acad. Sci. USA 91, 12,303–12,307.

    Article  CAS  Google Scholar 

  • Wolfgram F. and Myers L. (1973) Amyotrophic lateral sclerosis: effect of serum on anterior horn cells in tissue culture.Science 179, 579–580.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Research Institute of Biological Psychiatry, St. Hans Hospital, DK-4000, Roskilde, Denmark

    Ole Gredal, Michael Robin Witt, Kim Dekermendjian, Svend Erik Møller & Mogens Nielsen

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  1. Ole Gredal
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  2. Michael Robin Witt
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  3. Kim Dekermendjian
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  4. Svend Erik Møller
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  5. Mogens Nielsen
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Gredal, O., Witt, M.R., Dekermendjian, K. et al. Cerebrospinal fluid from amyotrophic lateral sclerosis has no effect on intracellular free calcium in cultured cortical neurons. Molecular and Chemical Neuropathology 29, 141–152 (1996). https://doi.org/10.1007/BF02814998

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  • DOI: https://doi.org/10.1007/BF02814998

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