Involvement of tubulin in MPP+ neurotoxicity on NGF‐differentiated PC12 cells.

In vivo, the neurotoxin MPTP is oxidated to MPP+, which is toxic to dopaminergic neurons. In this paper, we have used MPP+ as a tool to evoke neurotoxicity in the PC12 cell line and investigate the intracellular events that are involved. A cytotoxicity test, performed on undifferentiated and NGF‐differentiated PC12 cells, showed that MPP+ is much more toxic on differentiated cells and indicated the suitable range of concentrations for studying the starting events evoked by the neurotoxin. By indirect immunofluorescence we have shown that the localisation of α ‐ and β ‐tubulin in NGF‐differentiated cells was modified by a 24 h treatment with 15 μmol/l MPP+. A biochemical analysis was performed on cell extracts and the results showed that MPP+ treatment induced an increase in α ‐tubulin levels and a decrease in β ‐tubulin levels. These results suggest the involvement of the two microtubule proteins in MPP+ neurotoxicity on NGF‐differentiated PC12 cells.

[1]  Reaction of Lewy bodies with antibodies to phosphorylated and non-phosphorylated neurofilaments , 1986, Neuroscience Letters.

[2]  S. Orrenius,et al.  The mechanism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity: role of intracellular calcium. , 1988, Archives of biochemistry and biophysics.

[3]  M. Kirschner,et al.  Dynamic instability of microtubule growth , 1984, Nature.

[4]  D. Jacobowitz,et al.  A primate model of parkinsonism: selective destruction of dopaminergic neurons in the pars compacta of the substantia nigra by N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[5]  L. Greene,et al.  Release, storage and uptake of catecholamines by a clonal cell line of nerve growth factor (NGF) responsive pheochromocytoma cells , 1977, Brain Research.

[6]  L. Hartwell,et al.  Dominant effects of tubulin overexpression in Saccharomyces cerevisiae , 1989, Molecular and cellular biology.

[7]  M. Mattson Cellular signaling mechanisms common to the development and degeneration of neuroarchitecture. A review , 1989, Mechanisms of Ageing and Development.

[8]  J. Poirier,et al.  The specific vulnerability of the substantia nigra to MPTP is related to the presence of transition metals. , 1985, Biochemical and biophysical research communications.

[9]  B. Howard,et al.  A Dopaminergic Cell Line Variant Resistant to the Neurotoxin 1‐Methyl‐4‐Phenyl‐1,2,3,6‐Tetrahydropyridine , 1987, Journal of neurochemistry.

[10]  R. S. Burns,et al.  Intraneuronal generation of a pyridinium metabolite may cause drug-induced parkinsonism , 1984, Nature.

[11]  C. Mytilineou,et al.  1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine destroys dopamine neurons in explants of rat embryo mesencephalon. , 1984, Science.

[12]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[13]  W. Nicklas,et al.  Inhibition of NADH-linked oxidation in brain mitochondria by 1-methyl-4-phenyl-pyridine, a metabolite of the neurotoxin, 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine. , 1985, Life sciences.

[14]  R. Ramsay,et al.  Uptake of the neurotoxin 1-methyl-4-phenylpyridine (MPP+) by mitochondria and its relation to the inhibition of the mitochondrial oxidation of NAD+-linked substrates by MPP+. , 1986, Biochemical and biophysical research communications.

[15]  M. Bunge The axonal cytoskeleton: its role in generating and maintaining cell form , 1986, Trends in Neurosciences.

[16]  R. Duvoisin,et al.  Dopaminergic neurotoxicity of 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine in mice. , 1984, Science.

[17]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[18]  D. Cleveland Autoregulated control of tubulin synthesis in animal cells. , 1989, Current opinion in cell biology.

[19]  I. Grundke‐Iqbal,et al.  Lewy Bodies Contain Epitopes Both Shared and Distinct from Alzheimer Neurofibrillary Tangles , 1988, Journal of neuropathology and experimental neurology.

[20]  M. Camatini,et al.  N-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induces cytoskeletal alterations on ‘Swiss 3T3’ mouse fibroblasts , 1991, Neuroscience Letters.

[21]  L. Greene,et al.  Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M. Swash,et al.  Antibody to an abnormal protein in amyotrophic lateral sclerosis identifies Lewy body-like inclusions in ALS and Lewy bodies in Parkinson's disease , 1993, Neuroscience Letters.

[23]  D. Schubert,et al.  Storage and release of acetylcholine by a clonal cell line. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Snyder,et al.  Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6 -tetrahydropyridine: uptake of the metabolite N-methyl-4-phenylpyridine by dopamine neurons explains selective toxicity. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[25]  M. Zompo,et al.  Evaluation of the toxicity of the dopaminergic neurotoxins MPTP and MPP+ in PC12 pheochromocytoma cells: Binding and biological studies , 1988, Neuroscience Letters.