Proteomic analysis of differentiating neuroblastoma cells treated with sub-lethal neurite inhibitory concentrations of diazinon: Identification of novel biomarkers of effect
Autor: | John Flaskos, Alan J. Hargreaves, Wayne Harris, Magdalini Sachana |
---|---|
Rok vydání: | 2009 |
Předmět: |
Proteomics
Insecticides Neurite Cell Survival Cellular differentiation Blotting Western Fluorescent Antibody Technique Nerve Tissue Proteins macromolecular substances Biology Toxicology Microfilament Immunofluorescence Mice Neuroblastoma Differentiating Neuroblastoma Cell Line Tumor Neurites medicine Animals Electrophoresis Gel Two-Dimensional Cytoskeleton Pharmacology Dose-Response Relationship Drug medicine.diagnostic_test Cell Differentiation Cofilin Molecular biology Blot Actin Depolymerizing Factors Diazinon Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Cholinesterase Inhibitors Biomarkers Molecular Chaperones |
Zdroj: | Toxicology and Applied Pharmacology. 240:159-165 |
ISSN: | 0041-008X |
Popis: | In previous work we showed that sub-lethal levels of diazinon inhibited neurite outgrowth in differentiating N2a neuroblastoma cells. Western blotting analysis targeted at proteins involved in axon growth and stress responses, revealed that such exposure led to a reduction in the levels of neurofilament heavy chain, microtubule associated protein 1 B (MAP 1B) and HSP-70. The aim of this study was to apply the approach of 2 dimensional polyacrylamide gel electrophoresis and mass spectrometry to identify novel biomarkers of effect. A number of proteins were found to be up-regulated compared to the control on silver-stained gels. These were classified in to 3 main groups of proteins: cytosolic factors, chaperones and the actin-binding protein cofilin, all of which are involved in cell differentiation, survival or metabolism. The changes observed for cofilin were further confirmed by quantitative Western blotting analysis with anti-actin and anti-cofilin antibodies. Indirect immunofluorescence staining with the same antibodies indicated that the microfilament network was disrupted in diazinon-treated cells. Our data suggest that microfilament organisation is disrupted by diazinon exposure, which may be related to increased cofilin expression. |
Databáze: | OpenAIRE |
Externí odkaz: |