Neurotoxicity Research

, Volume 16, Issue 1, pp 77–86 | Cite as

Mechanism for Quinolinic Acid Cytotoxicity in Human Astrocytes and Neurons

  • Nady Braidy
  • Ross Grant
  • Seray Adams
  • Bruce J. Brew
  • Gilles J. GuilleminEmail author


There is growing evidence implicating the kynurenine pathway (KP) and particularly one of its metabolites, quinolinic acid (QUIN), as important contributors to neuroinflammation in several brain diseases. While QUIN has been shown to induce neuronal and astrocytic apoptosis, the exact mechanisms leading to cell death remain unclear. To determine the mechanism of QUIN-mediated excitotoxicity in human brain cells, we measured intracellular levels of nicotinamide adenine dinucleotide (NAD+) and poly(ADP-ribose) polymerase (PARP) and extracellular lactate dehydrogenase (LDH) activities in primary cultures of human neurons and astrocytes treated with QUIN. We found that QUIN acts as a substrate for NAD+ synthesis at very low concentrations (<50 nM) in both neurons and astrocytes, but is cytotoxic at sub-physiological concentrations (>150 nM) in both the cell types. We have shown that the NMDA ion channel blockers, MK801 and memantine, and the nitric oxide synthase (NOS) inhibitor, L-NAME, significantly attenuate QUIN-mediated PARP activation, NAD+ depletion, and LDH release in both neurons and astrocytes. An increased mRNA and protein expression of the inducible (iNOS) and neuronal (nNOS) forms of nitric oxide synthase was also observed following exposure of both cell types to QUIN. Taken together these results suggests that QUIN-induced cytotoxic effects on neurons and astrocytes are likely to be mediated by an over activation of an NMDA-like receptor with subsequent induction of NOS and excessive nitric oxide (NO)-mediated free radical damage. These results contribute significantly to our understanding of the pathophysiological mechanisms involved in QUIN neuro- and gliotoxicity and are relevant for the development of therapies for neuroinflammatory diseases.


Nitric oxide Quinolinic acid Astrocytes Neurons Alzheimer’s disease Neurodegeneration 


  1. Aguilera P, Chanez-Cardenas ME, Floriano-Snachez E, Barrera D, Santamaria A, Sanchez-Gonzalez DJ, Perez-Severiano F, Pedraza-Chaverri J, Jimenez PDM (2007) Time related changes in constitutive and inducible nitric oxide synthases in the rat striatum in a model of Huntington’s disease. Neurotoxicology 28:1200–1207PubMedCrossRefGoogle Scholar
  2. Atlante A, Gagliardi S, Minervini GM, Ciotti MT, Marra E, Calissano P (1997) Glutamate neurotoxicity in rat cerebellar granule cells: a major role of xanthine oxidase in oxygen radical formation. J Neurochem 68:2038–2045PubMedGoogle Scholar
  3. Ayata C, Ayata G, Hara H, Mathews RT, Beal MF, Ferrante RJ (1997) Mechanisms of reduced striatal NMDA excitotoxicity in type 1 nitric oxide synthase knock-out mice. J Neurosci 17:6908–6917PubMedGoogle Scholar
  4. Behan WM, McDonald M, Darlington LG, Stone TW (1999) Oxidative stress as a mechanism for quinolinic acid-induced hippocampal damage: protection by melatonin and deprenyl. Br J Pharmacol 128(8):1754–1760PubMedCrossRefGoogle Scholar
  5. Bernofsky C, Swan M (1973) An improved cycling assay for nicotinamide adenine dinucleotide. Anal Biochem 53:452–458PubMedCrossRefGoogle Scholar
  6. Bjorklund H, Eriksdotter-Nilsson M, Dahl D, Olson L (1984) Astrocytes in smears of CNS tissues as visualized by GFA and vimentin immunofluorescence. Med Biol 62(1):38–48PubMedGoogle Scholar
  7. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem 53:452–458Google Scholar
  8. Braidy N, Guillemin G, Grant R (2008) Promotion of cellular NAD+ anabolism: therapeutic potential for oxidative stress in ageing and Alzheimer’s disease. Neurotox Res 13(3, 4):173–184PubMedGoogle Scholar
  9. Cammer W (2002) Apoptosis of oligodendrocytes in secondary cultures from neonatal rat brains. Neurosci Lett 327(2):123–127PubMedCrossRefGoogle Scholar
  10. Conti F, DeBiasi S, Minelli A, Melone M (1996) Expression of NR1 and NR2A/B subunits of the NMDA receptor in cortical astrocytes. Glia 17(3):254–258PubMedCrossRefGoogle Scholar
  11. Dawson VL, Dawson TM, London ED, Bredt DS, Snyder SH (1991) Nitric oxide synthase mediates glutamate neurotoxicity in primary cortical cultures. Proc Natl Acad Sci USA 88:6368–6637PubMedCrossRefGoogle Scholar
  12. Dihne M, Block F, Korr H, Topper R (2001) Time course of glial proliferation and glial apoptosis following excitotoxic CNS injury. Brain Res 902(2):178–189PubMedCrossRefGoogle Scholar
  13. Du L, Zhang X, Han YY, Burke NA, Kochanek PM, Watkins SC, Grraham SH, Carcillo JA, Szabo C, Clark RS (2003) Intra-mitochondrial poly(ADP-ribosylation) contributes to NAD+ depletion and cell death induced by oxidative stress. J Biol Chem 278:18426–18433PubMedCrossRefGoogle Scholar
  14. Finkbeiner S, Cuero AM (2006) Disease modifying pathways in neurodegeneration. J Neurosci 26:10349–10357PubMedCrossRefGoogle Scholar
  15. Grant RS, Kapoor V (1998) Murine glial cells regenerate NAD, after peroxide-induced depletion, using either nicotinic acid, nicotinamide, or quinolinic acid as substrates. J Neurochem 70:1759–1763PubMedCrossRefGoogle Scholar
  16. Guillemin GJ, Brew BJ (2002) Implications of the kynurenine pathway and quinolinic acid in Alzheimer’s disease. Redox Rep 7(4):199–206PubMedCrossRefGoogle Scholar
  17. Guillemin GJ, Kerr SJ, Smythe GA, Smith DG, Kapoor V, Armati PJ, Croitoru J, Brew BJ (2001) Kynurenine pathway metabolism in human astrocytes: a paradox for neuronal protection. J Neurochem 78:1–13CrossRefGoogle Scholar
  18. Guillemin GJ, Meninger V, Brew BJ (2005a) Implications for the kynurenine pathway and quinolinic acid in amyotrophic lateral sclerosis. Neurodegener Dis 2(3–4):166–176PubMedCrossRefGoogle Scholar
  19. Guillemin GJ, Kerr SJ, Brew BJ (2005b) Involvement of quinolinic acid in AIDS dementia complex. Neurotox Res 7(1–2):103–123PubMedGoogle Scholar
  20. Guillemin GJ, Smythe G, Takikawa O, Brew BJ (2005c) Expression of indoleamine 2,3-dioxygenase and production of quinolinic acid by human microglia, astrocytes, and neurons. Glia 49(1):15–23PubMedCrossRefGoogle Scholar
  21. Guillemin GJ, Wang L, Brew BJ (2005d) Quinolinic acid selectively induces apoptosis of human astrocytes: potential role in AIDS dementia complex. J Neuroinflammation 2(16):1–6Google Scholar
  22. Guillemin GJ, Cullen KM, Lim CK, Smythe GA, Garner B, Kapoor V, Takikawa O, Brew BJ (2007) Characterization of the kynurenine pathway in human neurons. J Neurosci 27(47):12884–12892PubMedCrossRefGoogle Scholar
  23. Ha HC, Snyder SH (1999) Poly(ADP-ribose) polymerase is a mediator of necrotic cell death by ATP depletion. Proc Natl Acad Sci USA 96:13978–13982PubMedCrossRefGoogle Scholar
  24. Heyes MP (1993) Quinolinic acid and inflammation. Ann N Y Acad Sci 679:211–216PubMedCrossRefGoogle Scholar
  25. Heyes MP, Brew BJ, Martin A, Price RW, Salazar AM, Sidtis JJ, Yergey JA, Mouradian MM, Sadler AE, Keilp J et al (1991) Quinolinic acid in cerebrospinal fluid and serum in HIV-1 infection: relationship to clinical and neurological status. Ann Neurol 29(2):202–209PubMedCrossRefGoogle Scholar
  26. Heyes MP, Jordan EK, Lee K, Saito K, Frank JA, Snoy PJ, Markey SP, Gravell M (1992) Relationship of neurologic status in macaques infected with the simian immunodeficiency virus to cerebrospinal fluid quinolinic acid and kynurenic acid. Brain Res 570(1–2):237–250PubMedCrossRefGoogle Scholar
  27. Kelly WJ, Burke RE (1996) Apoptotic neuron death in rat substantia nigra induced by striatal excitotoxic injury is developmentally dependent. Neurosci Lett 220(2):85–88PubMedCrossRefGoogle Scholar
  28. Kerr SJ, Armati PJ, Brew BJ (1995) Neurocytotoxity of quinolinic acid in human brain cultures. J Neurovirol 1(5–6):375–380PubMedCrossRefGoogle Scholar
  29. Kerr SJ, Armati PJ, Guillemin GJ, Brew BJ (1998) Chronic exposure of human neurons to quinolinic acid results in neuronal changes consistent with AIDS dementia complex. AIDS 12(4):355–363PubMedCrossRefGoogle Scholar
  30. Koh JY, Choi DW (1987) Quantitative determination of glutamate mediated cortical neuronal injury in cell culture by lactate dehydrogenase efflux assay. J Neurosci Methods 20:83–90PubMedCrossRefGoogle Scholar
  31. Maldonado PD, Chanez-Cardenas ME, Barrera D, Villeda-Hernandez J, Santamaria A, Pedraza-Chaverri J (2007) Poly(ADP-ribose) polymerase-1 is involved in the neuronal death induced by quinolinic acid in rats. Neurosci Lett 425:28–33PubMedCrossRefGoogle Scholar
  32. Moroni F, Lombardi G, Moneti G, Aldinio C (1984) The excitotoxin quinolinic acid is present in the brain of several mammals and its cortical content increases during the aging process. Neurosci Lett 47(1):51–55PubMedCrossRefGoogle Scholar
  33. Perez-Severiano F, Escalante B, Rios C (1998) Nitric oxide synthase inhibition prevents acute quinolinate-induced striatal neurotoxicity. Neurochem Res 23:1297–1302PubMedCrossRefGoogle Scholar
  34. Possel H, Noack H, Putzke J, Wolf G, Seis H (2000) Selective upregulation of inducible nitric oxide synthase (iNOS) by lipopolysaccharide (LPS) and cytokines in microglia: in vitro and in vivo studies. Glia 32:51–59PubMedCrossRefGoogle Scholar
  35. Putt KS, Beilman GJ, Hergenrother PJ (2005) Direct quantification of poly(ADP-ribose) polymerase (PARP) activity as a means to distinguish necrotic and apoptotic death in cell and tissue samples. Chembiochem 6:53–55PubMedCrossRefGoogle Scholar
  36. Rya JK, Kim SU, McLarnon JG (2004) Blockade of quinolinic acid-induced neurotoxicity by pyruvate is associated with inhibition of glial activation in a model of Huntington’s disease. Exp Neurol 187:150–159CrossRefGoogle Scholar
  37. Santamaria A, Jimenez-Capdeville ME, Camacho A, Rodriguez-Martinez E, Flores A, Galvan-Arzate S (2001) In vivo hydroxyl radical formation after quinolinic acid infusion into rat corpus striatum. Neuroreport 12(12):2693–2696PubMedCrossRefGoogle Scholar
  38. Stone TW (1993) Neuropharmacology of quinolinic and kynurenic acids. Pharmacol Rev 45(3):309–379PubMedGoogle Scholar
  39. Stone TW (2001) Endogenous neurotoxins from tryptophan. Toxicon 39(1):61–73PubMedCrossRefGoogle Scholar
  40. Stone TW, Perkins MN (1981) Quinolinic acid: a potent endogenous excitant at amino acid receptors in CNS. Eur J Pharmacol 72(4):411–412PubMedCrossRefGoogle Scholar
  41. Ting KK, Brew B, Guillemin G (2007) The involvement of astrocytes and kynurenine pathway in Alzheimer’s disease. Neurotox Res 12(4):247–262PubMedCrossRefGoogle Scholar
  42. Velazquez JLP, Frantseva MV, Carlen PL (1997) In-vitro ischemia promotes glutamate-mediated free radical generation and intracellular calcium accumulation in hippocampal pyramidal neurones. J Neurosci 17:9085–9094Google Scholar
  43. Wong EHF, Kemp JA, Priestley T, Knight AR, Woodruff GN, Iversen LL (1986) The anticonvulsant MK-801 is a potent N-methyl-D-aspartate antagonist. Proc Natl Acad Sci USA 83:7104–7108PubMedCrossRefGoogle Scholar
  44. Zhang J, Dawson VL, Dawson TM, Snyder SH (1994) Nitric oxide activation of poly (ADP-ribose) synthetase in neurotoxicity. Science 263(5147):687–689PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Nady Braidy
    • 1
  • Ross Grant
    • 1
    • 2
  • Seray Adams
    • 1
  • Bruce J. Brew
    • 3
    • 4
  • Gilles J. Guillemin
    • 1
    • 3
    Email author
  1. 1.Department of Pharmacology, Faculty of MedicineUniversity of New South WalesSydneyAustralia
  2. 2.Australasian Research InstituteSydney Adventist HospitalSydneyAustralia
  3. 3.St Vincent’s Centre for Applied Medical ResearchSydneyAustralia
  4. 4.Department of NeurologySt Vincent’s HospitalSydneyAustralia

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