Calcium-dependent glutamate cytotoxicity in a neuronal cell line

Timothy H. Murphy, Alfred T. Malouf, Antonio Sastre, Ronald Lee Schnaar, Joseph T. Coyle

Research output: Contribution to journalArticle

Abstract

Membranes from the neuroblastoma × embryonic retina cell hybrid cell line, N18-RE-105, bind l-[3H]glutamate with a pharmacologic profile consistent with a 'quisqualate-type' brain l-glutamate receptor. We describe here the cytotoxic effect of l-glutamate receptor agonists on intact N18-RE-105 cells. Cytotoxicity was quantitated by measurement of the release of the cytosolic enzyme, lactate dehydrogenase, into the culture medium after addition of l-glutamate and its analogs to the cell culture medium. l-Glutamate (10 mM) and its confirmationally restricted analogs, quisqualate (1 mM) and ibotenate (10 mM), caused cell lysis. In contrast, similar analogs which do not bind to N18-RE-105 cell membranes (kainic acid, N-methyl-d,l-aspartic acid and γ-aminobutyric acid) were not cytotoxic. l-Glutamate-induced cytotoxicity was eliminated when calcium-free medium was used. Addition of inorganic or organic calcium channel antagonists also reduced the cytotoxicity of l-glutamate, even when 1.8 mM calcium was present in the medium. Cadmium chloride (10 μM) completely blocked l-glutamate toxicity, whereas manganese chloride (150 μM) and lanthanum chloride (25 μM) reduced toxicity by greater than 50%. Dihydropyridine voltage-sensitive calcium channel agonists or antagonists, had little or no significant effect on l-glutamate-induced toxicity. In contrast, the verapamil derivatives, D600 and D888, and the diltiazem derivative, MDL 12,330A reduced l-glutamate toxicity by greater than 50%. These results suggest that a subtype of voltage-sensitive calcium channels is involved in the mechanism of l-glutamate receptor mediated cytotoxicity in this cell line.

Original languageEnglish (US)
Pages (from-to)325-332
Number of pages8
JournalBrain Research
Volume444
Issue number2
DOIs
StatePublished - Mar 22 1988

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Glutamic Acid
Calcium
Cell Line
Glutamate Receptors
Quisqualic Acid
Calcium Channel Blockers
Culture Media
Calcium Channel Agonists
Gallopamil
Aminobutyrates
Excitatory Amino Acid Agonists
Cadmium Chloride
Diltiazem
Hybrid Cells
Kainic Acid
Calcium Channels
Verapamil
Neuroblastoma
L-Lactate Dehydrogenase
Aspartic Acid

Keywords

  • Calcium
  • Calcium channel
  • Cell culture
  • Excitotoxin
  • Glutamate
  • Neuroblastoma

ASJC Scopus subject areas

  • Developmental Biology
  • Molecular Biology
  • Clinical Neurology
  • Neuroscience(all)

Cite this

Calcium-dependent glutamate cytotoxicity in a neuronal cell line. / Murphy, Timothy H.; Malouf, Alfred T.; Sastre, Antonio; Schnaar, Ronald Lee; Coyle, Joseph T.

In: Brain Research, Vol. 444, No. 2, 22.03.1988, p. 325-332.

Research output: Contribution to journalArticle

Murphy, Timothy H. ; Malouf, Alfred T. ; Sastre, Antonio ; Schnaar, Ronald Lee ; Coyle, Joseph T. / Calcium-dependent glutamate cytotoxicity in a neuronal cell line. In: Brain Research. 1988 ; Vol. 444, No. 2. pp. 325-332.
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