Advertisement

Amino Acids

, Volume 9, Issue 2, pp 175–184 | Cite as

N-Acetyl-aspartylglutamate (NAAG) in human cerebrospinal fluid: Determination by high performance liquid chromatography, and influence of biological variables

  • V. Brovia
  • A. Ricciardi
  • L. Barbeito
Article
  • 49 Downloads

Summary

NAAG is one of the neuropeptides found in highest concentrations in the CNS. The presence of micromolar concentrations of NAAG in human CSF was demonstrated by using two different and complementary analytical approaches: 1) isocratic separation of endogenous NAAG by reverse-phase high performance liquid chromatography (HPLC) with dual wavelength detection and 2) derivatization of endogenous NAAG with acidic methanol and subsequent HPLC analysis of the derivative NAAG-trimethyl ester. The NAAG concentration was between 0.44µmol/l and 7.16µmol/l (mean of 2.19 ± 1.53µmol/l) in CSF samples from forty neuropsychiatric patients. Endogenous NAAG or [3H]NAAG added to CSF samples were not significantly degraded when the CSF was incubated at 37°C during one hour, suggesting that the peptide is a highly stable metabolite in the subarachnoid space. In addition, evidence is provided that NAAG does not present a concentration gradient along the lower subarachnoid space.

Keywords

Amino acids NAAG N-Acetyl-aspartyl-glutamate N-Acetyl aspartyl-glutamate trimethyl ester N-Acetyl-glutamyl-glutamate Cerebrospinal fluid-HPLC 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Blackely RD, Coyle JT (1988) The neurobiology of N-acetylaspartylglutamate. Int Rev Neurobiol 30: 39–100Google Scholar
  2. Blakely RD, Robinson MB, Thompson RC, Coyle JT (1988) Hydrolysis of the brain dipeptide N-acetyl-L-aspartyl-L-glutamate: subcellular and regional distribution, ontogeny, and the effect of lesions on N-acetylated-alpha-linked acidic dipeptidase activity. J Neurochem 50: 1200–1209Google Scholar
  3. Ferraro T, Hare TA (1985) Free and conjugated amino acids in human CSF: influence of age and sex. Brain Res 338: 53–60Google Scholar
  4. Grove J, Schechter PJ, Hanke NFJ, De Smet Y, Agid Y, Tell G, Koch-Weser J (1982) Concentration gradients of free and total aminobutyric acid and homocarnosine in human CSF: comparison of suboccipital and lumbar sampling. J Neurochem 39: 1618–1622Google Scholar
  5. Koller KJ, Zaczek R, Coyle JT (1984) N-acetyl-aspartyl-glutamate: regional levels in rat brain and the effects of brain lesions as determined by a new HPLC method. J Neurochem 43: 1136–1142Google Scholar
  6. Kornhuber J, Kornhuber ME, Hartmann GM, Kornhuber AW (1988)In vivo influences on cerebrospinal fluid amino acid levels. Neurochem Int 12: 25–31Google Scholar
  7. Luini A, Tal N, Goldberg O, Teichberg I (1984) An evaluation of selected brain constituents as putative excitatory neurotransmitters. Brain Res 324: 271–277Google Scholar
  8. Nyberg F, Nordström K, Terenius L (1985) Endopeptidase in human cerebrospinal fluid which cleaves proenkephalin B opioid peptides at consecutive basic amino acids. Biochem Biophys Res Commun 131: 1069–1074Google Scholar
  9. Nyberg F, Terenius L (1982) Endorphins in human cerebrospinal fluid. Life Sci 31: 1737–1740Google Scholar
  10. Ory-Lavollee L, Blakely RD, Coyle JT (1987) Neurochemical and immunocytochemical studies on the distribution of N-acetyl-aspartylglutamate and N-acetyl aspartate in rat spinal cord and some peripheral nervous tissues. J Neurochem 48: 895–899Google Scholar
  11. Pitkänen A, Matilainen R, Halonen T, Kutvonen R, Hartikainen P, Riekkinen P (1989) Inhibitory and excitatory amino acids in cerebrospinal fluid of chronic epileptic patients. J Neural Transm 76: 221–230Google Scholar
  12. Plaitakis A, Constantakakis E (1993) Altered metabolism of excitatory amino acids, N-acetyl-aspartate and N-acetyl-aspartyl-glutamate in amyotrophic lateral sclerosis. Brain Res Bull 30: 381–386Google Scholar
  13. Riveros N, Orrego F (1984) A study of possible excitatory effects of N-acetylaspartylglutamate in different in vivo and in vitro brain preparations. Brain Res 299: 393–395Google Scholar
  14. Rothstein JD, Tsai G, Kuncl RW, Clawson L, Cornblath DR, Drachman DB, Pestronk A, Stauch BL, Coyle JT (1990) Abnormal excitatory amino acid metabolism in amyotrophic lateral sclerosis. Ann Neurol 28: 18–25Google Scholar
  15. Serval V, Barbeito L, Pittaluga A, Chéramy A, Lavielle S, Glowinski J (1990) Competitive inhibition of N-acetylated-alpha-linked acidic dipeptidase activity by N-acetyl-L-aspartyl-beta-linked-L-glutamate. J Neurochem 55: 39–46Google Scholar
  16. Siciliano J, Laborde C, Barbeito L (1991) N-acetyl-aspartyl-glutamate in rat dorsal striatum: topographical distribution and effect of sensorimotor cortex lesions. Neurochem Int 19: 593–599Google Scholar
  17. Sjöström R, Ekstedt J, Änggard E (1975) Concentration gradients of monoamine metabolites in human cerebrospinal. J Neurol Neurosurg Psychiatry 38: 666–668Google Scholar
  18. Swahn C (1990) Determination of N-acetylaspartic acid in human cerebrospinal fluid by gas chromatography-mass spectrometry. J Neurochem 54: 1584–1588Google Scholar
  19. Tsai G, Stauch-Slusher B, Sim L, Hedreen JC, Rothstein JD, Kuncl R, Coyle JT (1991) Reductions in acidic amino acids and N-acetylaspartylglutamate in amyotrophic lateral sclerosis CNS. Brain Res 556: 151–156Google Scholar
  20. Wallasch M, Henning K, Lange U, Kuhn W, Eckhardt-Wallasch H (1988) Comparison of two methods for the determination of substance P immunoreactivity in human cerebrospinal fluid: sulphopropyl-Sephadex ion-exchange chromatography combined with radioimmunoassay. J Chromatogr 425: 175–181Google Scholar
  21. Whittemore ER, Koener JF (1989) An explanation for the purported excitation of piriform cortical neurons by N-acetyl-L-aspartyl-L-glutamic acid (NAAG). Proc Nat Acad Sci (USA) 86: 9602–9605Google Scholar
  22. Zollinger M, Amsler U, Brauchli J (1990) Quantification of N-acetylaspartylglutamate, an N-terminal blocked dipeptide neurotransmitter candidate, in brain slice superfusates by gas chromatography-mass spectrometry. J Chromatogr 532: 27–36Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • V. Brovia
    • 1
    • 2
  • A. Ricciardi
    • 1
  • L. Barbeito
    • 1
    • 2
  1. 1.División Neurobiología Celular y Molecular, Instituto Clemente EstableMontevideoUruguay
  2. 2.Sección Neurociencias, Facultad de CienciasUniversidad de la República MontevideoMontevideoUruguay

Personalised recommendations