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Polyamines pp 427-436 | Cite as

High-Resolution Capillary Gas Chromatography in Combination with Mass Spectrometry for Quantification of Three Major Polyamines in Postmortem Brain Cortex

  • Gary Gang Chen
  • Laura M. Fiori
  • Orval A. Mamer
  • Gustavo Turecki
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 720)

Abstract

There is considerable evidence supporting a role of the polyamine system in the etiology and pathology of mental disorders. Changes in the expression and activity of polyamine anabolic/catabolic enzymes, as well as in the levels of individual polyamines, have been found in many psychiatric conditions, including schizophrenia, mood disorders, anxiety, and suicidal behavior. Recent microarray studies have found that spermidine/spermine-N  1-acetyltransferase (SAT1, SSAT), the key enzyme in charge of the polyamine catabolic pathway, is downregulated in brain tissue of individuals who were depressed and died by suicide. To provide further insight into the downstream effects of altered SAT1 expression, we developed a quantitative gas chromatography-mass spectrometry method for measurement of polyamine concentrations in postmortem human brain tissues. This protocol employs a conventional electron ionization method with total ion and selected ion monitoring. This method can accurately measure the levels of the polyamines putrescine, spermidine, and spermine from very small quantities (1–50 mg) of postmortem brain tissues, with quantitation limits down to10 ng/g of wet tissue for putrescine and 100 ng/g for spermidine and spermine.

Key words

Polyamines Putrescine Putrescine-D4 Spermidine Spermine 1,7-Diaminoheptane Spermidine/spermine-N 1-acetyltransferase (SAT1, SSAT) Gas chromatography-mass spectrometry 

Notes

Acknowledgments

This work was supported by a Canadian Institutes for Health Research grant to GT (MOP-79253).

References

  1. 1.
    Casero RA, Pegg AE (2009) Polyamine catabolism and disease. Biochem J 421:323–338PubMedCrossRefGoogle Scholar
  2. 2.
    Igarashi K, Kashiwagi K (2010) Modulation of cellular function by polyamines. Int J Biochem Cell Biol 42:39–51 (originally published online 9 July 2009, at http://dx.doi.org/10.1016/j.biocel.2009.07.009)Google Scholar
  3. 3.
    Pegg AE (2009) Mammalian polyamine metabolism and function. IUBMB Life 61:880–894PubMedCrossRefGoogle Scholar
  4. 4.
    Casero RA Jr, Marton LJ (2007) Targeting polyamine metabolism and function in cancer and other hyperproliferative diseases. Nat Rev Drug Discov 6:373–390PubMedCrossRefGoogle Scholar
  5. 5.
    Pegg AE (2008) Spermidine/spermine-N(1)-acetyltransferase: a key metabolic regulator. Am J Physiol Endocrinol Metab 294:E995–E1010PubMedCrossRefGoogle Scholar
  6. 6.
    Sequeira A, Gwadry FG, Ffrench-Mullen JM, Canetti L, Gingras Y, Casero RA Jr et al (2006) Implication of SSAT by gene expression and genetic variation in suicide and major depression. Arch Gen Psychiatry 63:35–48PubMedCrossRefGoogle Scholar
  7. 7.
    Fiori LM, Turecki G (2008) Implication of the polyamine system in mental disorders. J Psychiatry Neurosci 33:102–110PubMedGoogle Scholar
  8. 8.
    Wallace HM, Fraser AV, Hughes A (2003) A perspective of polyamine metabolism. Biochem J 376:1–14PubMedCrossRefGoogle Scholar
  9. 9.
    Sequeira A, Klempan T, Canetti L, ffrench-Mullen J, Benkelfat C, Rouleau GA et al (2007) Patterns of gene expression in the limbic system of suicides with and without major depression. Mol Psychiatry 12:640–655PubMedCrossRefGoogle Scholar
  10. 10.
    Klempan TA, Rujescu D, Merette C, Himmelman C, Sequeira A, Canetti L et al (2009) Profiling brain expression of the spermidine/spermine N(1)-acetyltransferase 1 (SAT1) gene in suicide. Am J Med Genet B Neuropsychiatr Genet 150B:934–943PubMedCrossRefGoogle Scholar
  11. 11.
    Gilad GM, Gilad VH, Casanova MF, Casero RA Jr (1995) Polyamines and their metabolizing enzymes in human frontal cortex and hippocampus: preliminary measurements in affective disorders. Biol Psychiatry 38:227–234PubMedCrossRefGoogle Scholar
  12. 12.
    Wood PL, Khan MA, Moskal JR (2006) Neurochemical analysis of amino acids, polyamines and carboxylic acids: GC-MS quantitation of tBDMS derivatives using ammonia positive chemical ionization. J Chromatogr B Analyt Technol Biomed Life Sci 831:313–319PubMedCrossRefGoogle Scholar
  13. 13.
    Chen GG, Turecki G, Mamer OA (2009) A quantative GC-MS method for three major polyamines in post-mortem brain cortex. J Mass Spectrom 44:1203–1210PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Gary Gang Chen
    • 1
  • Laura M. Fiori
    • 1
  • Orval A. Mamer
    • 2
  • Gustavo Turecki
    • 3
  1. 1.McGill Group for Suicide StudiesDouglas Mental Health University Institute, McGill UniversityMontrealCanada
  2. 2.Mass Spectrometry UnitMcGill UniversityMontrealCanada
  3. 3.Douglas Mental Health University Institute, McGill UniversityMontrealCanada

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