Metabolism of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in a rat pheochromocytoma cell line, PC12h.
The uptake and metabolism of a neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were examined in a rat pheochromocytoma cell line, PC12h. These cells which contain only type A monoamine oxidase (MAO-A) oxidize MPTP into N-methyl-4-phenylpyridinium ion (MPP+). By kinetic analysis, the apparent Km value and the maximal velocity of the MPP+ production are 70.4 +/- 6.5 microM and 38.3 +/- 10.0 pmol/min/mg protein, respectively. After 7 days of culture in the presence of MPTP, the cells could oxidize from 25 to 50% of the MPTP added to the culture medium and could accumulate MPP+. The intracellular concentrations of MPTP were almost the same after 7 days of culture in the presence of MPTP from 10 nM to 100 microM. The cells could survive 7 days after exposure to up to 100 microM MPTP. Tyrosine hydroxylase (TH) and MAO activity were not affected by the presence of MPTP. Dopamine (DA) concentrations and a nonspecific enzyme, beta-galactosidase activity in the cells were not affected by the addition of MPTP. These data show that the uptake and oxidative conversion of MPTP take place in the cells having MAO-A alone, and that the neurotoxicity of MPP+ may not be due directly to its storage in subcellular compartments.
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D015655:1-Methyl-4-phenylpyridinium; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003010:Clorgyline / Q000494:pharmacology; D003470:Culture Media; D008995:Monoamine Oxidase / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D009994:Osmolar Concentration; D010673:Pheochromocytoma / Q000378:metabolism; D011725:Pyridines / Q000378:metabolism*; D011726:Pyridinium Compounds / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D051381:Rats; D012642:Selegiline / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D003470:Culture Media; D011725:Pyridines; D011726:Pyridinium Compounds; D012642:Selegiline; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D008995:Monoamine Oxidase; D003010:Clorgyline; D015655:1-Methyl-4-phenylpyridinium