Manganese-induced cytotoxicity in dopamine-producing cells

Denise L. Stredrick, Alan H. Stokes, Travis J. Worst, Willard M. Freeman, Elizabeth A. Johnson, Lawrence H. Lash, Michael Aschner, Kent E. Vrana

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Manganese (Mn) is an essential metal that, at excessive levels in the brain, produces extrapyramidal symptoms similar to those in patients with Parkinson's disease (PD). In the present study, Mn toxicity was characterized in a human neuroblastoma (SK-N-SH) cell line and in a mouse catecholaminergic (CATH.a) cell line. Mn was demonstrated to be more toxic in the catecholamine-producing CATH.a cells (EC50=60 μM) than in non-catecholaminergic SK-N-SH cells (EC50=200 μM). To test the hypothesis that the sensitivity of CATH.a cells to Mn is associated with their dopamine (DA) content, DA concentrations were suppressed in these cells by pretreatment with α-methyl-para-tyrosine (AMPT). Treatment for 24 h with 100 μM AMPT decreased intracellular DA, but offered no significant protection from Mn exposure (EC50=60 μM). Additional studies were carried out to assess if Mn toxicity was dependent on glutathione (GSH) levels. CATH.a cells were significantly protected by the addition of 5 mM GSH (Mn EC 50=200 μM) and 10 mM N-acetyl cysteine (NAC) (Mn EC 50=300 μM), therefore, indirectly identifying intracellular ROS formation as a mechanism for Mn neurotoxicity. Finally, apoptotic markers of Mn-induced cell death were investigated. DNA fragmentation, caspase-3 activation, and apoptosis-related gene expression were studied in CATH.a cells. No internucleosomal fragmentation or caspase activation was evident, even in the presence of "supraphysiological" Mn concentrations. cDNA hydridization array analysis with two differing Mn concentrations and time points, identified no noteworthy mRNA inductions of genes associated with programmed cell death. In conclusion, DA content was not responsible for the enhanced sensitivity of CATH.a cells to Mn toxicity, but oxidative stress was implicated as a probable mechanism of cytotoxicity.

Original languageEnglish (US)
Pages (from-to)543-553
Number of pages11
JournalNeuroToxicology
Volume25
Issue number4 SPEC. ISS.
DOIs
StatePublished - Jun 2004
Externally publishedYes

Fingerprint

Cytotoxicity
Manganese
Dopamine
Toxicity
Cell death
Cell Death
Acetylcysteine
Chemical activation
Cells
Cell Line
Oxidative stress
Poisons
DNA Fragmentation
Caspases
Oligonucleotide Array Sequence Analysis
Neuroblastoma
Gene expression
Caspase 3
Catecholamines
Glutathione

Keywords

  • CATH.a
  • Cytotoxicity
  • DNA fragmentation
  • Dopamine
  • Manganese

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Neuroscience(all)
  • Toxicology

Cite this

Stredrick, D. L., Stokes, A. H., Worst, T. J., Freeman, W. M., Johnson, E. A., Lash, L. H., ... Vrana, K. E. (2004). Manganese-induced cytotoxicity in dopamine-producing cells. NeuroToxicology, 25(4 SPEC. ISS.), 543-553. https://doi.org/10.1016/j.neuro.2003.08.006

Manganese-induced cytotoxicity in dopamine-producing cells. / Stredrick, Denise L.; Stokes, Alan H.; Worst, Travis J.; Freeman, Willard M.; Johnson, Elizabeth A.; Lash, Lawrence H.; Aschner, Michael; Vrana, Kent E.

In: NeuroToxicology, Vol. 25, No. 4 SPEC. ISS., 06.2004, p. 543-553.

Research output: Contribution to journalArticle

Stredrick, DL, Stokes, AH, Worst, TJ, Freeman, WM, Johnson, EA, Lash, LH, Aschner, M & Vrana, KE 2004, 'Manganese-induced cytotoxicity in dopamine-producing cells', NeuroToxicology, vol. 25, no. 4 SPEC. ISS., pp. 543-553. https://doi.org/10.1016/j.neuro.2003.08.006
Stredrick DL, Stokes AH, Worst TJ, Freeman WM, Johnson EA, Lash LH et al. Manganese-induced cytotoxicity in dopamine-producing cells. NeuroToxicology. 2004 Jun;25(4 SPEC. ISS.):543-553. https://doi.org/10.1016/j.neuro.2003.08.006
Stredrick, Denise L. ; Stokes, Alan H. ; Worst, Travis J. ; Freeman, Willard M. ; Johnson, Elizabeth A. ; Lash, Lawrence H. ; Aschner, Michael ; Vrana, Kent E. / Manganese-induced cytotoxicity in dopamine-producing cells. In: NeuroToxicology. 2004 ; Vol. 25, No. 4 SPEC. ISS. pp. 543-553.
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abstract = "Manganese (Mn) is an essential metal that, at excessive levels in the brain, produces extrapyramidal symptoms similar to those in patients with Parkinson's disease (PD). In the present study, Mn toxicity was characterized in a human neuroblastoma (SK-N-SH) cell line and in a mouse catecholaminergic (CATH.a) cell line. Mn was demonstrated to be more toxic in the catecholamine-producing CATH.a cells (EC50=60 μM) than in non-catecholaminergic SK-N-SH cells (EC50=200 μM). To test the hypothesis that the sensitivity of CATH.a cells to Mn is associated with their dopamine (DA) content, DA concentrations were suppressed in these cells by pretreatment with α-methyl-para-tyrosine (AMPT). Treatment for 24 h with 100 μM AMPT decreased intracellular DA, but offered no significant protection from Mn exposure (EC50=60 μM). Additional studies were carried out to assess if Mn toxicity was dependent on glutathione (GSH) levels. CATH.a cells were significantly protected by the addition of 5 mM GSH (Mn EC 50=200 μM) and 10 mM N-acetyl cysteine (NAC) (Mn EC 50=300 μM), therefore, indirectly identifying intracellular ROS formation as a mechanism for Mn neurotoxicity. Finally, apoptotic markers of Mn-induced cell death were investigated. DNA fragmentation, caspase-3 activation, and apoptosis-related gene expression were studied in CATH.a cells. No internucleosomal fragmentation or caspase activation was evident, even in the presence of {"}supraphysiological{"} Mn concentrations. cDNA hydridization array analysis with two differing Mn concentrations and time points, identified no noteworthy mRNA inductions of genes associated with programmed cell death. In conclusion, DA content was not responsible for the enhanced sensitivity of CATH.a cells to Mn toxicity, but oxidative stress was implicated as a probable mechanism of cytotoxicity.",
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