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Development of an in vitro model for cysteine dioxygenase expression in the brain

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Abstract

The development of an in vitro model for cysteine dioxygenase (CDO) expression in the brain would provide a useful model for determining the mechanisms for the regulation of CDO expression that does not involve the use of animals. Here we demonstrate the screening and characterization of a cell line that expresses CDO, the primary metabolizing enzyme of cysteine and the regulatory point of sulfate production. A panel of four commercially available tumor-derived human brain cell lines, each representing one major class of brain cell, were screened using western blotting and activity assay for cysteine dioxygenase expression. One cell line, TE 671 (human medulloblastoma) was found to express both a protein of approximately 70 kDa and CDO activity. Nuclease protection assay (NPA) of mRNA isolated from TE 671 showed the expression of a CDO mRNA. Reverse transcription–polymerase chain reaction of this mRNA and sequencing of the cDNA obtained showed that this was indeed CDO. Treatment of TE 671 cells with cysteine resulted in the upregulation of CDO mRNA, whereas treatment with tumor necrosis factor α resulted in the downregulation of CDO mRNA, as evidenced using NPA. The characterization of an in vitro model for CDO expression provides a useful tool for the investigation of this important enzyme, which may have an etiological role in the pathogenesis of Parkinson's disease.

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References

  • Adams JD, Odunze IN. Oxygen free radicals and Parkinson's disease. Free Radic Biol Med. 1991;10:161-9.

    Article  PubMed  CAS  Google Scholar 

  • Andersen ME, Meister A. Marked increase of cysteine levels in many regions of the brain after administration of 2-oxothiazolidine-4-carboxylate. FASEB J. 1989;3:1632-6.

    Google Scholar 

  • Akerlund B, Jarstrand C, Lindeke B, Sonnerborg A, Akerblad AC, Rasool O. Effect of N-acetylcysteine (NAC) treatment on HIV-1 infection: a double-blind placebo-controlled trial. Eur J Clin Pharmacol. 1996;50:457-61.

    Article  PubMed  CAS  Google Scholar 

  • Beal MF. Excitotoxicity and nitric oxide in Parkinson's disease pathogenesis. Ann Neurol. 1998;44:S110-14.

    Article  PubMed  CAS  Google Scholar 

  • Bella DL, Hahn C, Stipanuk MH. Effects of non-sulfur and sulfur amino acids on the regulation of hepatic enzymes of cysteine metabolism. Am J Physiol. 1999;277:E144-53.

    PubMed  CAS  Google Scholar 

  • Boss V, Nutt KM, Conn PJ. L-Cysteinesulphinic acid as an endogenous agonist of a novel metabotropic receptor coupled to stimulation of phospholipase D activity. Mol Pharmacol. 1994;45:1177-82.

    PubMed  CAS  Google Scholar 

  • Bradford MM. A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem. 1976;72: 248-54.

    Article  PubMed  CAS  Google Scholar 

  • Callans DJ, Wacker LS, Mitchell MC. Effects of ethanol feeding and withdrawal on plasma glutathione elimination in the rat. Hepatology. 1987;7:496-501.

    PubMed  CAS  Google Scholar 

  • Daniels KM, Stipanuk MH. The effect of dietary cysteine levels upon cysteine metabolism in rats. J Nutr. 1982;112:2130-41.

    PubMed  CAS  Google Scholar 

  • De Bandt JP, Lim SK, Plassart F, et al. Independent and combined actions of interleukin-?, tumour necrosis factor a and glucagon on amino acid metabolism in the isolated perfused rat liver. Metab Clin Exp. 1994;43:822-9.

    PubMed  CAS  Google Scholar 

  • Dexter DT, Sian J, Rose S, et al. Indices of oxidative stress and mitochondrial function in individuals with incidental Lewy body disease. Ann Neurol. 1994;35:38-44.

    Article  PubMed  CAS  Google Scholar 

  • Doble A. The role of excitotoxicity in neurodegenerative disease: implications for therapy. Pharmacol Ther. 1999;81:163-221.

    Article  PubMed  CAS  Google Scholar 

  • Gaitonde MK. A spectrophotometric method for the direct determination of cysteine in the presence of other naturally occurring amino acids. Biochem J. 1967;104:627-33.

    PubMed  CAS  Google Scholar 

  • Gerlach M, Riederer P. Animal models of Parkinson's disease: an empirical comparison with the phenomenology of the disease in man. J Neural Transm. 1996;103:987-1041.

    Article  PubMed  CAS  Google Scholar 

  • Hosokawa Y, Yamaguchi K, Kohashi N, Kori Y, Fujii O, Ueda I. Immaturity of the enzyme activity and the response to inducers of rat liver cysteine dioxygenase during rat devel-opment. J Biochem. 1980;88:389-94.

    PubMed  CAS  Google Scholar 

  • Hosokawa Y, Matsumoto A, Oka J, Itakura H, Yamaguchi K. Isolation and characterisation of a cDNA for rat liver cysteine dioxygenase. Biochem Biophys Res Commun. 1990;168:471-8.

    Article  Google Scholar 

  • Ida S, Okhuma S, Kimori M, Kuriyama K, Morimoto N, Ibata Y. Regulatory role of cysteine dioxgyenase in cerebral biosynthesis of taurine. Analysis using cerebellum from 3-acetylpyridine treated rats. Brain Res. 1985;344(1):62-9.

    Article  PubMed  CAS  Google Scholar 

  • Jeremiah S, McCann KP, Williams AC, et al. Chromosomal localisation of genes encoding for human and mouse liver cytosolic cysteine dioxygenase. Ann Hum Gen. 1996;60:29-33.

    CAS  Google Scholar 

  • Kohashi N, Yamaguchi K, Hosokawa Y, Kori Y, Fuji O, Ueda I. Dietary control of cysteine dioxygenase in rat liver. J Biochem. 1978;84:159-68.

    PubMed  CAS  Google Scholar 

  • Laemmli, UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680-5.

    Article  PubMed  CAS  Google Scholar 

  • Li H, Dryhurst G. Irreversible inhibition of mitochondiral complex I by 7-(2-aminoethyl)-3,4-dihydro-5-hydroxy-2H-1,4-benzothiazine-3-carboxylic acid (DHBT-1): a putatuve nigral endotoxin relevant to Parkinson's disease. J Neuro-chem. 1997;69:1530-41.

    CAS  Google Scholar 

  • McCann KP, Akbari MT, Williams AC, Ramsden DB. Human cysteine dioxygenase type-I: primary structure derived from base sequencing of cDNA. Biochim Biophys Acta. 1994; 1209:107-10.

    PubMed  CAS  Google Scholar 

  • Misra CH. Effect of morphine on cysteine oxidase activity in the rat brain and reproductive tracts of rats. Biochem Pharmacol. 1979;28(10):1695-8.

    Article  PubMed  CAS  Google Scholar 

  • Nunn PB, Davis AJ, O'Brien P. Carbamate formation and the neurotoxicity of L-?-amino acids. Science. 1991;251:1619.

    PubMed  CAS  Google Scholar 

  • Olanow CW. An introduction to the free radical hypothesis of Parkinson's disease. Ann Neurol. 1992;32:S2-9.

    Article  PubMed  CAS  Google Scholar 

  • Olney JW, Ho OL. Brain damage in infant mice following oral intake of glutamate, aspartate or cysteine. Nature. 1970;227:609-10.

    Article  PubMed  CAS  Google Scholar 

  • Olney JW, Zorumski CF, Price MT, Labruyere JL. L-Cysteine, a bicarbonate-sensitive excitotoxin. Science. 1990a;248:596-9.

    PubMed  CAS  Google Scholar 

  • Olney JW, Zorumski CF, Stewart GR, Price MT, Wang GJ, Labruyere J. Excitotoxicity of L-DOPA and 6-OH-DOPA: implications for Parkinson's and Huntington's diseases. Exp Neurol. 1990b;108:269-72.

    Article  PubMed  CAS  Google Scholar 

  • Ono H, Sakamoto A, Eguchi T, et al. Plasma total homocysteine concentrations in epileptic patients taking anti-convulsants. Metab Clin Exp. 1997;46:959-62.

    PubMed  CAS  Google Scholar 

  • Ott L, McClain CJ, Gillespie M, Young B. Cytokines and metabolic dysfunction after severe head injury. J Neurotrauma. 1994;11:447-72.

    Article  PubMed  CAS  Google Scholar 

  • Parsons RB, Waring RH, Ramsden DB, Williams AC. Toxicity of cysteine and cysteinesulphinic acid to human neuronal cell-lines. J Neurol Sci. 1997;152(supplement):S62-6.

    Article  PubMed  CAS  Google Scholar 

  • Parsons RB, Ramsden DB, Waring RH, Barber PC, Williams AC. Hepatic localisation of rat cysteine dioxygenase. J Hepatol. 1998a;29:595-602.

    Article  PubMed  CAS  Google Scholar 

  • Parsons RB, Barber PC, Waring RH, Williams AC, Ramsden DB. Cysteine dioxygenase: regional expression of activity in rat brain. Neurosci Lett. 1998b;248:101-4.

    Article  PubMed  CAS  Google Scholar 

  • Pean AR, Parsons RB, Waring RH, Williams AC, Ramsden DB. Toxicity of sulphur-containing compounds to neuronal cell-lines. J Neurol Sci. 1995;19(supplement):107-8.

    Article  Google Scholar 

  • Perry TL, Norman MG, Yong VW, et al. Hallervorden-Spatz disease: cysteine accumulation and cysteine dioxygenase deficiency in the globul pallidus. Ann Neurol. 1985;18:482-9.

    Article  PubMed  CAS  Google Scholar 

  • Potgieter HC, Ubbink JB, Bissbort S, Bester MJ, Spies JH, Vermaak WJ. Spontaneous exidation of methionine: effect on the quantification of plasma methionine levels. Anal Biochem. 1997;248:86-93.

    Article  PubMed  CAS  Google Scholar 

  • Rerat A, Nunes CS, Mendy F, Roger L. Amino acid absorption and production of pancreatic hormones in non-anaesthetized pigs after duodenal infusions of a milk enzymic hydrosylate or of free amino acids. Br J Nutr. 1988;60:121-36.

    Article  PubMed  CAS  Google Scholar 

  • Sakakibara S, Yamaguchi K, Hosokawa Y, Kohashi N, Ueda I, Sakamoto Y. Purification and some properties of rat liver cysteine oxidase (cysteine dioxygenase). Biochim Biophys Acta. 1976;422:273-9.

    PubMed  CAS  Google Scholar 

  • Schapira AHV, Copper JM, Dexter D, Jenner P, Clark JB, Marsden CD. Mitochondrial complex-I deficiency in Parkinson's disease. Lancet. 1989;I (8649):1269.

    Article  Google Scholar 

  • Seaton TA, Cooper JM, Schapira AH. Free radical scavengers protect dopaminergic cell lines from apoptosis induced by complex I inhibitors. Brain Res. 1997;777:110-8.

    Article  PubMed  CAS  Google Scholar 

  • Shattuck KE, Grinnel CD, Goldman AS, Rassin DK. The acute effects of TNF-? on the isolated perfused rat liver. J Invest Med. 1996;44(2):64-9.

    CAS  Google Scholar 

  • Sörbo B, Ewetz L. The enzymic oxidation of cysteine to cysteinesulphinate in rat liver. Biochem Biophys Res Commun. 1965;3:359-63.

    Article  Google Scholar 

  • Steventon GB, Heafield MTE, Waring RH, Williams AC. Drug metabolism in Parkinson's disease. Neurology. 1989;39: 883-7.

    PubMed  CAS  Google Scholar 

  • Tayek JA. Effects of tumour necrosis factor ? on skeletal muscle amino acid metabolism studied in vivo. J Am College Nutr. 1996;15:164-8.

    CAS  Google Scholar 

  • Varastet M, Riche D, Maziere M, Hantraye P. Chronic MPTP treatment reproduces in baboons the differential vulnerability of mesencephalic dopaminergic neurones observed in Parkinson's disease. Euroscience. 1994;63:47-56.

    Article  CAS  Google Scholar 

  • Van Guldener C, Janssen MJ, de Meer K, Donker AJ, Stehouwer CD. Effect of folic acid and betaine on fasting and postmethionine-loading plasma homocysteine and methionine levels in chronic haemodialysis patients. J Intern Med. 1999;245:175-83.

    Article  PubMed  CAS  Google Scholar 

  • Waelsch H, Rittenberg D. Glutathione I. The metabolism of glutathione studied with isotopic glycine. J Biol Chem. 1941;139:671-774.

    Google Scholar 

  • Warren RS, Starnes HF Jr, Gabrilove JL, Oettgen HF, Brennan MF. The acute metabolic effects of tumour necrosis factor administration in humans. Arch Surg. 1987;122:1396-400.

    PubMed  CAS  Google Scholar 

  • Wilcken DE, Dudman NP, Tyrrell PA. Homocystinuria due to cystathionine beta-synthase deficiency-the effects of betaine treatment in pyridoxine-responsive patients. Metab Clin Exp. 1985;34:1115-21.

    PubMed  CAS  Google Scholar 

  • Yamaguchi K, Hosokawa Y. The stabilising protein, protein-A, of cysteine dioxygenase. Adv Exp Med Biol. 1987;217:29-38.

    PubMed  CAS  Google Scholar 

  • Yamaguchi K, Sakakibara S, Koga K, Ueda I. Induction and activation of cysteine oxidase in rat liver I. The effects of cysteine, hydrocortisone and nicotinamide injections on hepatic cysteine oxidase and tyrosine transaminase activities of intact and adrenalectomised rats. Biochim Biophys Acta. 1971;237:502-12.

    CAS  Google Scholar 

  • Yamaguchi K, HosokawaY, Kohashi N, Kori Y, Sakakibara S, Ueda I. Rat liver cysteine dioxygenase (cysteine oxidase). Further purification, characterisation and analysis of the activation and inactivation. J Biochem. 1978;83:479-91.

    PubMed  CAS  Google Scholar 

  • Yasmineh WG, Tsai MY, Theologides A. Hepatic mitochondrial enzyme activity and serum amino acid composition in rats treated with tumour necrosis factor. Life Sci. 1995;56: 621-7.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. School of Biosciences, Queen Elizabeth Hospital, University of Birmingham, Edgbaston, Birmingham, B15 2TH, UK

    S. Qusti & R.H. Waring

  2. Department of Medicine, Queen Elizabeth Hospital, University of Birmingham, Edgbaston, Birmingham, B15 2TH, UK

    R.B. Parsons, K.D.H. Abouglila & D.B. Ramsden

  3. Centre for Neurosciences, Queen Elizabeth Hospital, University of Birmingham, Edgbaston, Birmingham, B15 2TH, UK

    A.C. Williams

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  1. S. Qusti
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  6. D.B. Ramsden
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Qusti, S., Parsons, R., Abouglila, K. et al. Development of an in vitro model for cysteine dioxygenase expression in the brain. Cell Biol Toxicol 16, 243–255 (2000). https://doi.org/10.1023/A:1007634110983

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