Neurochemical Research

, Volume 36, Issue 3, pp 443–451 | Cite as

Non-invasive Monitoring of L-2-Oxothiazolidine-4-Carboxylate Metabolism in the Rat Brain by In vivo 13C Magnetic Resonance Spectroscopy

  • Michael P. Gamcsik
  • M. Daniel Clark
  • Susan M. Ludeman
  • James B. Springer
  • Michael A. D’Alessandro
  • Nicholas E. Simpson
  • Roxana Pourdeyhimi
  • C. Bryce Johnson
  • Stephanie D. Teeter
  • Stephen J. Blackband
  • Peter E. Thelwall
Original Paper


The cysteine precursor L-2-oxothiazolidine-4-carboxylate (OTZ, procysteine) can raise cysteine concentration, and thus glutathione levels, in some tissues. OTZ has therefore been proposed as a prodrug for combating oxidative stress. We have synthesized stable isotope labeled OTZ (i.e. L-2-oxo-[5-13C]-thiazolidine-4-carboxylate, 13C-OTZ) and tracked its uptake and metabolism in vivo in rat brain by 13C magnetic resonance spectroscopy. Although uptake and clearance of 13C-OTZ was detectable in rat brain following a bolus dose by in vivo spectroscopy, no incorporation of isotope label into brain glutathione was detectable. Continuous infusion of 13C-OTZ over 20 h, however, resulted in 13C-label incorporation into glutathione, taurine, hypotaurine and lactate at levels sufficient for detection by in vivo magnetic resonance spectroscopy. Examination of brain tissue extracts by mass spectrometry confirmed only low levels of isotope incorporation into glutathione in rats treated with a bolus dose and much higher levels after 20 h of continuous infusion. In contrast to some previous studies, bolus administration of OTZ did not alter brain glutathione levels. Even a continuous infusion of OTZ over 20 h failed to raise brain glutathione levels. These studies demonstrate the utility of in vivo magnetic resonance for non-invasive monitoring of antioxidant uptake and metabolism in intact brain. These types of experiments can be used to evaluate the efficacy of various interventions for maintenance of brain glutathione.


Magnetic resonance Metabolism Cysteine Glutathione Taurine 


  1. 1.
    Schulz JB, Lindenau J, Seyfried J et al (2000) Glutathione, oxidative stress and neurodegeneration. Eur J Biochem 267:4904–4911CrossRefPubMedGoogle Scholar
  2. 2.
    Osman LP, Mitchell SC, Waring RH (1997) Cysteine, its metabolism and toxicity. Sulfur Rep 20:155–172CrossRefGoogle Scholar
  3. 3.
    Meister A (1989) A brief history of glutathione and a survey of its metabolism and functions. In: Dolphin D, Poulson R, Avramovic O (eds) Glutathione. Chemical biochemical and medical aspects, part A. Wiley, New York, pp 1–48Google Scholar
  4. 4.
    Heard KJ (2008) Acetylcysteine for acetaminophen poisoning. N Engl J Med 359:285–289CrossRefPubMedGoogle Scholar
  5. 5.
    Williamson JM, Meister A (1981) Stimulation of hepatic glutathione formation by administration of L-2-oxothiazolidine-4-carboxylate, a 5-oxo-L-prolinase Substrate. Proc Natl Acad Sci USA 78:936–939CrossRefPubMedGoogle Scholar
  6. 6.
    Arakawa M, Ito Y (2007) N-acetylcysteine and neurodegenerative diseases: basic and clinical pharmacology. Cerebellum 6:308–314CrossRefGoogle Scholar
  7. 7.
    Arfsten DP, Johnson EW, Wilfong ER et al (2007) Distribution of radio-labeled N-acetyl-L-cysteine in Sprague-Dawley rats and its effect on glutathione metabolism following single and repeat dosing by oral gavage. Cutan Ocul Toxicol 26:113–134CrossRefPubMedGoogle Scholar
  8. 8.
    Cudkowicz ME, Sexton ME, Ellis T et al (1999) The pharmacokinetics and pharmaco-dynamics of Procysteine in amyotrophic lateral sclerosis. Neurology 52:1492–1494PubMedGoogle Scholar
  9. 9.
    Anderson ME, Meister A (1989) Marked increase of cysteine levels in many regions of the brain after administration of 2-oxothiazolidine-4-carboxylate. FASEB J 3:1632–1636PubMedGoogle Scholar
  10. 10.
    Mesina JE, Page RH, Hetzel FW et al (1989) Administration of L-2-oxothiazolidine-4-carboxylate increases glutathione levels in rat brain. Brain Res 478:181–183CrossRefPubMedGoogle Scholar
  11. 11.
    Park SW, Kim SH, Park KH et al (2004) Preventive effects of antioxidants in MPTP-induced mouse model of Parkinson’s disease. Neurosci Lett 363:243–246CrossRefPubMedGoogle Scholar
  12. 12.
    Thelwall PE, Yemin AY, Gillian TL et al (2005) Noninvasive in vivo detection of glutathione metabolism in tumors. Cancer Res 65:10149–10153CrossRefPubMedGoogle Scholar
  13. 13.
    Berk M, Ng F, Dean O et al (2008) Glutathione; a novel treatment target in psychiatry. Trends Pharmacol Sci 29:346–351CrossRefPubMedGoogle Scholar
  14. 14.
    Zeevalk GD, Razmpour R, Bernard LP (2008) Glutathione and Parkinson’s disease: is this the elephant in the room? Biomed Pharmacother 62:236–249CrossRefPubMedGoogle Scholar
  15. 15.
    Amoyaw PNA, Springer JB, Gamcsik MP et al (2010) Synthesis of 13C-labelled derivatives of cysteine for magnetic resonance imaging studies of drug uptake and conversion to glutathione in rat brain. J Labelled Compds Radiopharm (in press)Google Scholar
  16. 16.
    Millis KK, Lesko SA, Gamcsik MP (1997) Formation, intracellular distribution and efflux of glutathione-bimane conjugates in drug-sensitive and -resistant MCF-7 cells. Cancer Chemother Pharmacol 40:101–111CrossRefPubMedGoogle Scholar
  17. 17.
    Boogers I, Plugge W, Stokkermans YQ et al (2008) Ultra-performance liquid chromatographic analysis of amino acid in protein hydrolysates using an automated pre-column derivatisation method. J Chromatogr A 1189:406–409CrossRefPubMedGoogle Scholar
  18. 18.
    Cohen SA, DeAntonis K, Michaud DP (1993) Compositional protein analysis using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, a novel derivatization reagent. In: Angeletti RH (eds) Techniques in protein chemistry IV. Academic Press, Inc, San Diego, pp 289–298Google Scholar
  19. 19.
    Manura JJ, Manura DJ (2009) Isotope distribution calculator and mass spec plotter.
  20. 20.
    Gerard-Monnier D, Fougeat S, Gourvest JF et al (1993) Partial prevention of glutathione depletion in rats following acute intoxication with diethylmaleate. Clin Physiol Biochem 10:36–42PubMedGoogle Scholar
  21. 21.
    Shivakumar BR, Ravindranath V (1992) Selective modulation of glutathione in mouse brain regions and its effect on acrylamide-induced neurotoxicity. Biochem Pharmacol 43:263–269CrossRefPubMedGoogle Scholar
  22. 22.
    Pileblad E, Magnusson T (1992) Increase in rat brain glutathione following intracerebroventricular administration of γ-glutamylcysteine. Biochem Pharmacol 44:895–903CrossRefPubMedGoogle Scholar
  23. 23.
    Reed MC, Thomas RL, Pavisic J et al (2008) A mathematical model of glutathione metabolism. Theor Biol Med Modeling 5:8Google Scholar
  24. 24.
    Choi I-Y, Gruetter R (2004) Dynamic or inert metabolism? Turnover of N-acetyl aspartate and glutathione from D-[1–13C]glucose in the rat brain in vivo. J Neurochem 91:778–787CrossRefPubMedGoogle Scholar
  25. 25.
    Dominy JE Jr, Hwang J, Stipanuk MH (2007) Overexpression of cysteine dioxygenase reduces intracellular cysteine and glutathione pools in HepG2/C3A cells. Am J Physiol Endocrinol Metab 293:E62–E69CrossRefPubMedGoogle Scholar
  26. 26.
    Stipanuk MH, Dominy JE Jr, Lee JI et al (2006) Mammalian cysteine metabolism: new insights into regulation and cysteine metabolism. J Nutr 136:1652S–1659SPubMedGoogle Scholar
  27. 27.
    Beetsch JW, Olson JE (1998) Taurine synthesis and cysteine metabolism in cultured rat astrocytes: effects of hyperosmotic exposure. Am J Physiol (Cell Physiol) 274:C866–C874Google Scholar
  28. 28.
    Stipanuk MH, Londono M, Lee JI et al (2002) Enzymes and metabolites and cysteine metabolism in nonhepatic tissues of rats show little response to changes in dietary protein or sulfur amino acid levels. J Nutr 132:3369–3378PubMedGoogle Scholar
  29. 29.
    Dominy J, Eller S, Dawson R Jr (2004) Building biosynthetic schools: Reviewing compartmentation of CNS taurine synthesis. Neurochem Res 29:97–103CrossRefPubMedGoogle Scholar
  30. 30.
    Pasantes-Morales H, Chatagner F, Mandel P (1980) Synthesis of taurine in rat liver and brain in vivo. Neurochem Res 5:441–451CrossRefPubMedGoogle Scholar
  31. 31.
    Stipanuk MH, Ueki I (2010) Dealing with methionine/homocysteine sulfur: cysteine metabolism to taurine and inorganic sulfur. J Inherit Metab Dis. doi:10.1007/s10545-009-9006-9
  32. 32.
    Shibuya N, Tanaka M, Yoshida M et al (2009) 3-Mercaptopyruvate sulfurtransferase produces hydrogen sulfide and bound sulfane sulfur in the brain. Antioxid Redox Signal 11:703–714CrossRefPubMedGoogle Scholar
  33. 33.
    Potter DW, Tran TB (1993) Apparent rates of glutathione turnover in rat tissues. Toxicol Appl Pharmacol 120:186–192CrossRefPubMedGoogle Scholar
  34. 34.
    Brand A, Leibfritz D, Hamprecht B et al (1998) Metabolism of cysteine in astroglial cells: synthesis of hypotaurine and taurine. J Neurochem 71:827–832CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Michael P. Gamcsik
    • 1
  • M. Daniel Clark
    • 2
  • Susan M. Ludeman
    • 3
  • James B. Springer
    • 4
  • Michael A. D’Alessandro
    • 3
  • Nicholas E. Simpson
    • 5
  • Roxana Pourdeyhimi
    • 1
  • C. Bryce Johnson
    • 1
  • Stephanie D. Teeter
    • 1
  • Stephen J. Blackband
    • 2
  • Peter E. Thelwall
    • 6
  1. 1.UNC/NCSU Joint Department of Biomedical EngineeringRaleighUSA
  2. 2.Department of Neuroscience, McKnight Brain InstituteUniversity of FloridaGainesvilleUSA
  3. 3.Departments of Arts and Sciences and Pharmaceutical SciencesAlbany College of Pharmacy and Health SciencesAlbanyUSA
  4. 4.Department of MedicineDuke University Medical CenterDurhamUSA
  5. 5.Department of MedicineUniversity of FloridaGainesvilleUSA
  6. 6.Newcastle Magnetic Resonance Centre, Campus for Ageing and VitalityNewcastle UniversityNewcastle upon TyneUK

Personalised recommendations