Requirement of a dopaminergic neuronal phenotype for toxicity of low concentrations of 1-methyl-4-phenylpyridinium to human cells

Journal Article · Sun Nov 15 00:00:00 EST 2009 · Toxicology and Applied Pharmacology

DOI:https://doi.org/10.1016/j.taap.2009.07.027· OSTI ID:21344785

Schildknecht, Stefan ^[1]; Poeltl, Dominik; Nagel, Daniel M; Matt, Florian; Scholz, Diana; Lotharius, Julie; Schmieg, Nathalie; Salvo-Vargas, Alberto; Leist, Marcel ^[1]

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1. University of Konstanz, Konstanz (Germany)

LUHMES cells are conditionally-immortalized non-transformed human fetal cells that can be differentiated to acquire a dopaminergic neuron-like phenotype under appropriate growth conditions. After differentiation by GDNF and cyclic adenosine monophosphate, LUHMES were sensitive to 1-methyl-4-phenylpyridinium (MPP{sup +}) toxicity at <= 5 muM, but resistant to the parental compound 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The high homogeneity and purity of the cultures allowed the detection of metabolic changes during the degeneration. Cellular ATP dropped in two phases after 24 and 48 h; cellular glutathione (GSH) decreased continuously, paralleled by an increase in lipid peroxidation. These events were accompanied by a time-dependent degeneration of neurites. Block of the dopamine transporter by GBR 12909 or mazindol completely abrogated MPP{sup +} toxicity. Inhibition of de novo dopamine synthesis by alpha-methyl-L-tyrosine or 3-iodo-L-tyrosine attenuated toxicity, but did not reduce the initial drop in ATP. Inhibition of mixed lineage kinases by CEP1347 completely prevented the MPP{sup +}-induced loss of viability and intracellular GSH, but failed to attenuate the initial drop of ATP. For the quantitative assessment of neurite degeneration, an automated imaging-based high content screening approach was applied and confirmed the findings made by pharmacological interventions in this study. Our data indicate that inhibition of mitochondrial ATP synthesis is not sufficient to trigger cell death in MPP{sup +}-treated LUHMES.

• OSTI ID: 21344785
• Journal Information: Toxicology and Applied Pharmacology, Vol. 241, Issue 1; Other Information: DOI: 10.1016/j.taap.2009.07.027; PII: S0041-008X(09)00313-5; Copyright (c) 2009 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; ISSN 0041-008X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
APOPTOSIS
DOPAMINE
GLUTATHIONE
MEN
MITOCHONDRIA
NERVE CELLS
NERVOUS SYSTEM DISEASES
PHENOTYPE
PYRIDINIUM COMPOUNDS
SYNTHESIS
TOXICITY
TYROSINE
AMINES
AMINO ACIDS
AMMONIUM COMPOUNDS
ANIMAL CELLS
ANIMALS
AROMATICS
AUTONOMIC NERVOUS SYSTEM AGENTS
AZINES
CARBOXYLIC ACIDS
CARDIOTONICS
CARDIOVASCULAR AGENTS
CELL CONSTITUENTS
DISEASES
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROXY ACIDS
HYDROXY COMPOUNDS
MALES
MAMMALS
MAN
NEUROREGULATORS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PEPTIDES
PHENOLS
POLYPEPTIDES
POLYPHENOLS
PRIMATES
PROTEINS
PYRIDINES
QUATERNARY AMMONIUM COMPOUNDS
RADIOPROTECTIVE SUBSTANCES
RESPONSE MODIFYING FACTORS
SOMATIC CELLS
SYMPATHOMIMETICS
VERTEBRATES