There is a need for efficient methods giving more insight into complex mechanisms of neurotoxicity. The uses of in vitro test methods and testing strategies have been proposed to comply with this requirement. With the present study we aimed to develop a novel in vitro approach which mimics the in vivo complexity and detects comprehensively neurotoxicity. For this purpose we combined rat primary re-aggregating brain cell cultures with a mass spectrometry (MS) based metabolomics approach. For the proof of principle we treated cultures for 48 hours with the neurotoxicant methyl mercury chloride (0.1 100 µM) and the brain stimulant caffeine (1-100 µM). Cell lysate samples were prepared and their cellular metabolic profiles were obtained and analyzed by principal component analyses (PCA). The results showed PCA plots with concentration dependent cluster formations at non-cytotoxic concentrations of methyl mercury chloride (0.1-1 µM) and caffeine (1-100 µM). Moreover, significant metabolic perturbations were revealed and considered as putative biomarkers for their neurotoxic effects. Using MS fragmentation analyses the most prominent putative biomarkers were identified to be -aminobutyric acid, choline, glutamine, creatine and spermine. The quantification of their spectral mass ion intensities demonstrated concentration dependent alterations which were in agreement with previous studies and hypothesized mechanisms of neurotoxicity and neuroprotection. In addition, we evaluated whether the approach could identify neurotoxicity by testing eight compounds at a non-cytototoxic concentrations having target organ toxicity for liver, kidney or brain. The results showed a PCA plot with cluster formations largely dependent on target organ toxicity indicating the potential of the approach to identify the neurotoxicity of compounds. Overall, results propose the performance of a validation study to determine its relevance and applicability for neurotoxicity screening. In conclusion, for the first time we show the benefits and utility of in vitro metabolomics to detect comprehensively neurotoxicity and to discover new biomarkers.

VAN VLIET Erwin;  MORATH Siegfried;  ESKES Chantra;  LINGE Jens;  RAPPSILBER Juri;  HONEGGER Paul;  HARTUNG Thomas;  COECKE Sandra; 

2008-02-07

ELSEVIER SCIENCE BV

JRC37499

0161-813X,   

https://publications.jrc.ec.europa.eu/repository/handle/JRC37499,   

10.1016/j.neuro.2007.09.007,