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Diazoxon Disrupts the Expression and Distribution of β III ‐Tubulin and MAP 1B in Differentiating N2a Cells
Author(s) -
Sachana Magdalini,
Sidiropoulou Erasmia,
Flaskos John,
Harris Wayne,
Robinson Alex J.,
Woldehiwet Zerai,
Hargreaves Alan J.
Publication year - 2014
Publication title -
basic and clinical pharmacology and toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.805
H-Index - 90
eISSN - 1742-7843
pISSN - 1742-7835
DOI - 10.1111/bcpt.12192
Subject(s) - tubulin , microtubule , neurite , cytoskeleton , neurofilament , immunofluorescence , microtubule associated protein , monoclonal antibody , neurotoxicity , microbiology and biotechnology , biology , isotype , chemistry , cell , biochemistry , antibody , in vitro , immunohistochemistry , immunology , toxicity , organic chemistry
This study aimed at assessing the effects of diazoxon ( DZO ), a major metabolite of the insecticide diazinon ( DZ ), on key cytoskeletal proteins in differentiating N2a neuroblastoma cells. Initial experiments established that sublethal concentrations of 1, 5 and 10 μM DZO produced profound inhibition of neurite outgrowth. Densitometric scanning of probed immunoblots of N2a cell lysates demonstrated that DZO had no effect on total β‐tubulin levels. However, probing with a monoclonal antibody that recognised specifically the β III ‐tubulin isotype revealed that 10 μM DZO induced a significant reduction in the levels of this particular form. Levels of polyglutamylated tubulin were not altered. Exposure to 10 μM DZO also decreased the expression of microtubule‐associated protein 1B ( MAP 1B). However, DZO had no effect on the expression of MAP tau. DZO also failed to affect the levels neurofilament light ( NFL ) and neurofilament medium ( NFM ) chain levels. Indirect immunofluorescence demonstrated that the staining of neurites in treated cells was weaker than in the controls for β III ‐tubulin. In conclusion, DZO disrupts the microtubule ( MT ) network affecting the expression and distribution of two specific MT proteins known to be important in neuritogenesis. DZO may contribute to the developmental neurotoxicity seen following exposure to DZ .