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Diazinon oxon interferes with differentiation of rat C6 glioma cells
E Sidiropoulou1, M Sachana, J Flaskos
1Department of Veterinary Pathology, Faculty of Veterinary Science, University of Liverpool, Leahurst, CH64 7TE, UK.
Abstract:
The purpose of this study was to evaluate the toxicity of diazinon oxon (DZO), a major in vivo metabolite of the organophosphate insecticide diazinon (DZ), on differentiating rat C6 glioma cells. At concentrations shown to be non-cytotoxic by both the MTT and the Kenacid blue dye binding assays (1, 5 and 10 microM), DZO caused after 24h a reduction in the number of extensions developed from C6 cells induced to differentiate by serum withdrawal and addition of sodium butyrate. Densitometric scanning of Western blots of extracts of C6 cells demonstrated that, at all concentrations used, DZO decreased after 24h the expression of glial fibrillary acidic protein (GFAP) compared to controls. In addition, exposure to 10 microM DZO for 24h reduced the levels of tubulin and microtubule associated protein 1B (MAP1B). On the other hand, levels of MAP2c were not affected by DZO treatment. In contrast to our previous data on DZ, the above findings suggest that its oxon metabolite, DZO, may, at biologically relevant, subcytotoxic concentrations, interfere with glial cell differentiation.
Insights
Diazinon oxon (DZO), a metabolite of the insecticide diazinon, can disrupt glial cell differentiation. Even at low, non-cytotoxic doses, DZO reduces cell extensions and key protein expression in rat C6 glioma cells.
Area of Science:
- Neuroscience
- Toxicology
- Cell Biology
Background:
- Organophosphate insecticides like diazinon (DZ) are widely used.
- Diazinon oxon (DZO) is a primary active metabolite of diazinon.
- Understanding the specific toxicity of DZO on neural cells is crucial.
Purpose of the Study:
- To investigate the toxicity of diazinon oxon (DZO) on differentiating rat C6 glioma cells.
- To determine if sub-cytotoxic concentrations of DZO affect glial cell differentiation markers.
Main Methods:
- Rat C6 glioma cells were induced to differentiate using serum withdrawal and sodium butyrate.
- Cell viability was assessed using MTT and Kenacid blue dye binding assays.
- DZO toxicity was evaluated at non-cytotoxic concentrations (1, 5, 10 microM).
- Changes in cell morphology (extensions) were quantified.
- Protein expression levels (GFAP, tubulin, MAP1B, MAP2c) were analyzed using Western blotting.
Main Results:
- Non-cytotoxic concentrations of DZO (1, 5, 10 microM) reduced the number of cell extensions in differentiating C6 cells after 24 hours.
- DZO significantly decreased the expression of glial fibrillary acidic protein (GFAP) at all tested concentrations.
- Exposure to 10 microM DZO reduced levels of tubulin and microtubule-associated protein 1B (MAP1B).
- MAP2c levels remained unaffected by DZO treatment.
Conclusions:
- Diazinon oxon (DZO), at biologically relevant, sub-cytotoxic concentrations, interferes with glial cell differentiation.
- DZO's effects on differentiation markers like GFAP suggest a specific mechanism of neurotoxicity distinct from its parent compound, diazinon.
- These findings highlight the potential risks of DZO exposure to neural development and function.
