An in vitro model for toxicological investigations of environmental neurotoxins in primary neuronal cell cultures.
Currently, most neurotoxicological investigations are still conducted using various animal models (e.g. chickens, rodents). In this report, alternative strategies of testing were examined to detect the neurotoxic potency of foreign compounds. Primary neuronal cell cultures from fetal rats are already an accepted model for mechanistic and pharmacological studies in drug research. Their suitability for neurotoxicological studies was examined by using industrial model compounds, which are well-known inductors of neuropathies: acrylamide, hexachlorophene, paraquat, n-hexane, and its neurotoxic metabolites acetylaceton and 2,5-hexandione. As a control compound, the nonneurotoxic solvent n-heptane was used. General cytotoxicity and the intracellular content of glial fibrillary acid protein, neuron-specific enolase, and neurofilaments were measured. n-Heptane induced an acute cytotoxicity and acrylamide and 2,5-hexandione produced a delayed cytotoxicity in primary neuronal cells, whereas the others showed no cytotoxic potency in the tested concentration range. These results were in agreement with the quantification of neurons by neuron-specific enolase. In contrast, with the exception of acetylaceton, glia cells were significantly affected by all neurotoxins at the later time. Signs of axonopathies were demonstrated for acrylamide, n-hexane and its metabolites, as well as for hexachlorophene and paraquat in vitro, by determining the intracellular neurofilament level. Therefore, the determination of cell-specific end points is necessary to detect the neurotoxic potency and quality of a compound, whereas the cytotoxicity assay limited the tested concentration range.
Schmuck
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BAYER Pharma Research Center, Wuppertal, Germany.
Schlüter
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D020106:Acrylamide; D000178:Acrylamides / Q000633:toxicity; D000704:Analysis of Variance; D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology* / Q000187:drug effects / Q000196:embryology; D003432:Cross-Linking Reagents / Q000633:toxicity; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006540:Herbicides / Q000633:toxicity*; D006582:Hexachlorophene / Q000633:toxicity; D006586:Hexanes / Q000378:metabolism / Q000633:toxicity; D006588:Hexanones / Q000633:toxicity; D007205:Indium Radioisotopes; D007382:Intermediate Filaments / Q000187:drug effects; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D009498:Neurotoxins / Q000633:toxicity*; D010269:Paraquat / Q000633:toxicity; D010422:Pentanones / Q000633:toxicity; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar
D000178:Acrylamides; D003432:Cross-Linking Reagents; D006540:Herbicides; D006586:Hexanes; D006588:Hexanones; D007205:Indium Radioisotopes; D009498:Neurotoxins; D010422:Pentanones; D020106:Acrylamide; C026385:n-hexane; C008790:acetylacetone; C011269:2,5-hexanedione; D010751:Phosphopyruvate Hydratase; D006582:Hexachlorophene; D010269:Paraquat