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Effects of 1‐Methyl‐4‐Phenyl‐1,2,3,6‐Tetrahydropyridine on Differentiating Mouse N2a Neuroblastoma Cells
Author(s) -
De Girolamo Luigi A.,
Billett E. Ellen,
Hargreaves Alan J.
Publication year - 2000
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.2000.0750133.x
Subject(s) - mptp , neurotoxin , neurotoxicity , neurofilament , blot , microbiology and biotechnology , cell culture , biology , phosphorylation , chemistry , axon , toxicity , endocrinology , biochemistry , immunology , immunohistochemistry , dopamine , genetics , dopaminergic , organic chemistry , gene
: The effect of the neurotoxin 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) was investigated in mouse N2a neuroblastoma cells, induced to differentiate by serum withdrawal and addition of dibutyryl cyclic AMP, over a 24‐h period. Addition of MPTP (10 μ M ) during differentiation caused a change in cell morphology characterised by an inhibition of axon outgrowth, in the absence of cell death. Biochemical characterisation by western blotting revealed that MPTP had no significant effects on the levels of actin, α‐tubulin, or total heavy‐chain neurofilament (NF‐H). However, NF‐H phosphorylation appeared to increase following MPTP treatment when blots were probed with the phosphorylation state‐specific antibodies RMd09 and Ta51. In addition, indirect immunofluorescence analysis revealed an accumulation of phosphorylated NF‐H in the cell perikaryon, suggesting that altered NF‐H distribution was associated with the observed effects of MPTP on cell morphology. These changes may represent a useful in vitro marker of MPTP neurotoxicity within a simple differentiating neuronal cell model system.