Dopamine toxicity in neuroblastoma cells: Role of glutathione depletion by L-BSO and apoptosis

A. H. Stokes, D. Y. Lewis, L. H. Lash, W. G. Jerome, K. W. Grant, Michael Aschner, K. E. Vrana

Research output: Contribution to journalArticle

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Abstract

Dopamine (DA), while an essential neurotransmitter, is also a known neurotoxin that potentially plays an etiologic role in several neurodegenerative diseases. DA metabolism and oxidation readily produce reactive oxygen species (ROS) and DA can also be oxidized to a reactive quinone via spontaneous, enzyme-catalyzed or metal-enhanced reactions. A number of these reactions are cytotoxic, yet the precise mechanisms by which DA leads to cell death remain unknown. In this study, the neuroblastoma cell line, SK-N-SH, was utilized to examine DA toxicity under varying oxidant states. Cells pretreated with the glutathione (GSH)-depleting compound, L-buthionine sulfoximine (L-BSO), exhibited enhanced sensitivity to DA compared to controls (non-GSH-depleted cells). Furthermore, in cells pretreated with L-BSO, the addition of ascorbate (250 μM) afforded significant protection against DA-induced toxicity, while pyruvate (500 μM) had no protective effect. To further characterize the possibility that DA is associated with oxidative stress, additional studies were carried out with manganese (30 μM) as a pro-oxidant. Manganese and DA (200 μM), although not cytotoxic when individually administered to SK-N-SH cells, had a synergistic action on cytotoxicity. Finally, morphological and molecular markers of programmed cell death (apoptosis) were observed in cells treated with DA and L-BSO. These markers included membrane blebbing and internucleosomal DNA fragmentation. These results suggest that DA toxicity is tightly linked to intracellular oxidant/antioxidant levels, and that environmental factors, such as excessive Mn exposure, may modulate cellular sensitivity to DA. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)1-8
Number of pages8
JournalBrain Research
Volume858
Issue number1
DOIs
StatePublished - 2000
Externally publishedYes

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Buthionine Sulfoximine
Neuroblastoma
Glutathione
Dopamine
Apoptosis
Manganese
Oxidants
Reactive Oxygen Species
Cell Death
Neurotoxins
DNA Fragmentation
Blister
Pyruvic Acid
Neurodegenerative Diseases
Neurotransmitter Agents

Keywords

  • Apoptosis
  • Dopamine
  • Glutathione
  • Manganese

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Stokes, A. H., Lewis, D. Y., Lash, L. H., Jerome, W. G., Grant, K. W., Aschner, M., & Vrana, K. E. (2000). Dopamine toxicity in neuroblastoma cells: Role of glutathione depletion by L-BSO and apoptosis. Brain Research, 858(1), 1-8. https://doi.org/10.1016/S0006-8993(99)02329-X

Dopamine toxicity in neuroblastoma cells : Role of glutathione depletion by L-BSO and apoptosis. / Stokes, A. H.; Lewis, D. Y.; Lash, L. H.; Jerome, W. G.; Grant, K. W.; Aschner, Michael; Vrana, K. E.

In: Brain Research, Vol. 858, No. 1, 2000, p. 1-8.

Research output: Contribution to journalArticle

Stokes, A. H. ; Lewis, D. Y. ; Lash, L. H. ; Jerome, W. G. ; Grant, K. W. ; Aschner, Michael ; Vrana, K. E. / Dopamine toxicity in neuroblastoma cells : Role of glutathione depletion by L-BSO and apoptosis. In: Brain Research. 2000 ; Vol. 858, No. 1. pp. 1-8.
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AB - Dopamine (DA), while an essential neurotransmitter, is also a known neurotoxin that potentially plays an etiologic role in several neurodegenerative diseases. DA metabolism and oxidation readily produce reactive oxygen species (ROS) and DA can also be oxidized to a reactive quinone via spontaneous, enzyme-catalyzed or metal-enhanced reactions. A number of these reactions are cytotoxic, yet the precise mechanisms by which DA leads to cell death remain unknown. In this study, the neuroblastoma cell line, SK-N-SH, was utilized to examine DA toxicity under varying oxidant states. Cells pretreated with the glutathione (GSH)-depleting compound, L-buthionine sulfoximine (L-BSO), exhibited enhanced sensitivity to DA compared to controls (non-GSH-depleted cells). Furthermore, in cells pretreated with L-BSO, the addition of ascorbate (250 μM) afforded significant protection against DA-induced toxicity, while pyruvate (500 μM) had no protective effect. To further characterize the possibility that DA is associated with oxidative stress, additional studies were carried out with manganese (30 μM) as a pro-oxidant. Manganese and DA (200 μM), although not cytotoxic when individually administered to SK-N-SH cells, had a synergistic action on cytotoxicity. Finally, morphological and molecular markers of programmed cell death (apoptosis) were observed in cells treated with DA and L-BSO. These markers included membrane blebbing and internucleosomal DNA fragmentation. These results suggest that DA toxicity is tightly linked to intracellular oxidant/antioxidant levels, and that environmental factors, such as excessive Mn exposure, may modulate cellular sensitivity to DA. Copyright (C) 2000 Elsevier Science B.V.

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