title: Characterization of the kynurenine pathway in NSC-34 cell line: implications for amyotrophic lateral sclerosis.
abstract: Amyotrophic lateral sclerosis (ALS) is the most common type of motor neuron degenerative disease for which the aetiology is still unknown. The kynurenine pathway (KP) is a major degradative pathway of tryptophan ultimately leading to the production of NAD(+) and is also one of the major regulatory mechanisms of the immune response. The KP is known to be involved in several neuroinflammatory disorders. Among the KP intermediates, quinolinic acid (QUIN) is a potent excitotoxin, while kynurenic acid and picolinic acid are both neuroprotectant. This study aimed to (i) characterize the components of the KP in NSC-34 cells (a rodent motor neuron cell line) and (ii) assess the effects of QUIN on the same cells. RT-PCR and immunocytochemistry were used to characterize the KP enzymes, and lactate dehydrogenase (LDH) test was used to assess the effect of QUIN in the absence and presence of NMDA receptor antagonists, kynurenines and 1-methyl tryptophan. Our data demonstrate that a functional KP is present in NSC-34 cells. LDH tests showed that (i) QUIN toxicity on NSC-34 cells increases with time and concentration; (ii) NMDA antagonists, 2-amino-5-phosphonopentanoic acid, MK-801 and memantine, can partially decrease QUIN toxicity; (iii) kynurenic acid can decrease LDH release in a linear manner, whereas picolinic acid does the same but non-linearly; and (iv) 1-methyl tryptophan is effective in decreasing QUIN release by the rodent microglial cell line BV-2 and thus protects NSC-34 from cell death. There is currently a lack of effective treatment for ALS and our in vitro results provide a novel therapeutic strategy for ALS patients.
authors.affiliation: Department of Pharmacology, School of Medical Sciences, University of New South Wales, Sydney, New South Wales, Australia.
authors.forename: Yiquan
authors.initials: Y
authors.lastname: Chen
authors.forename: Bruce J
authors.initials: BJ
authors.lastname: Brew
authors.forename: Gilles J
authors.initials: GJ
authors.lastname: Guillemin
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D006867:Hydrolases / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology; D007502:Iron Chelating Agents / Q000494:pharmacology; D007737:Kynurenine / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D017628:Microglia / Q000187:drug effects / Q000378:metabolism; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism*; D010848:Picolinic Acids / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors; D014364:Tryptophan / Q000031:analogs & derivatives / Q000494:pharmacology; D014366:Tryptophan Oxygenase / Q000378:metabolism
chemical_list: D007502:Iron Chelating Agents; D010848:Picolinic Acids; D007737:Kynurenine; C028129:tryptophan methyl ester; D007371:Interferon-gamma; D014364:Tryptophan; D007770:L-Lactate Dehydrogenase; D014366:Tryptophan Oxygenase; D006867:Hydrolases; C021480:kynureninase; C030614:picolinic acid