Journal of Inherited Metabolic Disease

, Volume 33, Issue 2, pp 113–120 | Cite as

Oligodendrocyte development and myelinogenesis are not impaired by high concentrations of phenylalanine or its metabolites

  • Renaud Schoemans
  • Marie-Stéphane Aigrot
  • Chaohong Wu
  • Raphaël Marée
  • Pengyu Hong
  • Shibeshi Belachew
  • Claire Josse
  • Catherine Lubetzki
  • Vincent Bours
Original Article


Phenylketonuria (PKU) is a metabolic genetic disease characterized by deficient phenylalanine hydroxylase (PAH) enzymatic activity. Brain hypomyelination has been reported in untreated patients, but its mechanism remains unclear. We therefore investigated the influence of phenylalanine (Phe), phenylpyruvate (PP), and phenylacetate (PA) on oligodendrocytes. We fisrt showed in a mouse model of PKU that the number of oligodendrocytes is not different in corpus callosum sections from adult mutants or from control brains. Then, using enriched oligodendroglial cultures, we detected no cytotoxic effect of high concentrations of Phe, PP, or PA. Finally, we analyzed the impact of Phe, PP, and PA on the myelination process in myelinating cocultures using both an in vitro index of myelination, based on activation of the myelin basic protein (MBP) promoter, and the direct quantification of myelin sheaths by both optical measurement and a bioinformatics method. None of these parameters was affected by the increased levels of Phe or its derivatives. Taken together, our data demonstrate that high levels of Phe, such as in PKU, are unlikely to directly induce brain hypomyelination, suggesting involvement of alternative mechanisms in this myelination defect.


Corpus Callosum Myelin Basic Protein Myelin Sheath Large Neutral Amino Acid Oligodendrocyte Development 

List of abbreviations




Phenylalanine hydroxylase










Large neutral amino acids


Oligodendrocyte progenitor cells


Subventricular zone



The authors thank the Imaging Platform, the SPF Animal Facility, and the Neurosciences Department from GIGA-Research for their experimental help. We also thank Olympus Belgium N.V. for providing the Cell^R system.

Competing interest

None declared.


  1. Ahring K, Belanger-Quintana A, Dokoupil K et al (2009) Dietary management practices in phenylketonuria across European centres. Clin Nutr 28(3):231–236CrossRefPubMedGoogle Scholar
  2. Barbin G, Aigrot MS, Charles P et al (2004) Axonal cell-adhesion molecule L1 in CNS myelination. Neuron Glia Biol 1(1):65–72CrossRefPubMedGoogle Scholar
  3. Baumann N, Pham-Dinh D (2001) Biology of oligodendrocyte and myelin in the mammalian central nervous system. Physiol Rev 81(2):871–927PubMedGoogle Scholar
  4. Bottenstein J, Hayashi I, Hutchings S et al (1979) The growth of cells in serum-free hormone-supplemented media. Methods Enzymol 58:94–109CrossRefPubMedGoogle Scholar
  5. Cabib S, Pascucci T, Ventura R, Romano V, Puglisi-Allegra S (2003) The behavioral profile of severe mental retardation in a genetic mouse model of phenylketonuria. Behav Genet 33(3):301–310CrossRefPubMedGoogle Scholar
  6. Charles P, Hernandez MP, Stankoff B et al (2000) Negative regulation of central nervous system myelination by polysialylated-neural cell adhesion molecule. Proc Natl Acad Sci U S A 97(13):7585–7590CrossRefPubMedGoogle Scholar
  7. Demerens C, Stankoff B, Zalc B, Lubetzki C (1999) Eliprodil stimulates CNS myelination: new prospects for multiple sclerosis? Neurology 52(2):346–350PubMedGoogle Scholar
  8. Dyer CA (2000) Comments on the neuropathology of phenylketonuria. Eur J Pediatr 159(Suppl 2):S107–S108CrossRefPubMedGoogle Scholar
  9. Dyer CA, Kendler A, Philibotte T, Gardiner P, Cruz J, Levy HL (1996) Evidence for central nervous system glial cell plasticity in phenylketonuria. J Neuropathol Exp Neurol 55(7):795–814CrossRefPubMedGoogle Scholar
  10. Gow A, Friedrich VL Jr, Lazzarini RA (1992) Myelin basic protein gene contains separate enhancers for oligodendrocyte and Schwann cell expression. J Cell Biol 119(3):605–616CrossRefPubMedGoogle Scholar
  11. Hoeksma M, Reijngoud DJ, Pruim J, de Valk HW, Paans AM, van Spronsen FJ (2009) Phenylketonuria: high plasma phenylalanine decreases cerebral protein synthesis. Mol Genet Metab 96(4):177–182CrossRefPubMedGoogle Scholar
  12. Horster F, Schwab MA, Sauer SW et al (2006) Phenylalanine reduces synaptic density in mixed cortical cultures from mice. Pediatr Res 59(4 Pt 1):544–548CrossRefPubMedGoogle Scholar
  13. Hughes JV, Johnson TC (1976) The effects of phenylalanine on amino acid metabolism and protein synthesis in brain cells in vitro. J Neurochem 26(6):1105–1113CrossRefPubMedGoogle Scholar
  14. Huttenlocher PR (2000) The neuropathology of phenylketonuria: human and animal studies. Eur J Pediatr 159(Suppl 2):S102–S106CrossRefPubMedGoogle Scholar
  15. Joseph B, Dyer CA (2003) Relationship between myelin production and dopamine synthesis in the PKU mouse brain. J Neurochem 86(3):615–626CrossRefPubMedGoogle Scholar
  16. Kaufman S (1989) An evaluation of the possible neurotoxicity of metabolites of phenylalanine. J Pediatr 114(5):895–900CrossRefPubMedGoogle Scholar
  17. Matalon R, Michals-Matalon K, Bhatia G et al (2007) Double blind placebo control trial of large neutral amino acids in treatment of PKU: effect on blood phenylalanine. J Inherit Metab Dis 30(2):153–158CrossRefPubMedGoogle Scholar
  18. McCarthy KD, de Vellis J (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85(3):890–902CrossRefPubMedGoogle Scholar
  19. McDonald JD, Charlton CK (1997) Characterization of mutations at the mouse phenylalanine hydroxylase locus. Genomics 39(3):402–405CrossRefPubMedGoogle Scholar
  20. Miller RH (2002) Regulation of oligodendrocyte development in the vertebrate CNS. Prog Neurobiol 67(6):451–467CrossRefPubMedGoogle Scholar
  21. Pardridge WM (1998) Blood-brain barrier carrier-mediated transport and brain metabolism of amino acids. Neurochem Res 23(5):635–644CrossRefPubMedGoogle Scholar
  22. Pascucci T, Ventura R, Puglisi-Allegra S, Cabib S (2002) Deficits in brain serotonin synthesis in a genetic mouse model of phenylketonuria. NeuroReport 13(18):2561–2564CrossRefPubMedGoogle Scholar
  23. Pey AL, Desviat LR, Gamez A, Ugarte M, Perez B (2003) Phenylketonuria: genotype-phenotype correlations based on expression analysis of structural and functional mutations in PAH. Hum Mutat 21(4):370–378CrossRefPubMedGoogle Scholar
  24. Rocha JC, Martel F (2009) Large neutral amino acids supplementation in phenylketonuric patients. J Inherit Metab Dis 32(4):472–480CrossRefPubMedGoogle Scholar
  25. Sarkissian CN, Boulais DM, McDonald JD, Scriver CR (2000a) A heteroallelic mutant mouse model: a new orthologue for human hyperphenylalaninemia. Mol Genet Metab 69(3):188–194CrossRefPubMedGoogle Scholar
  26. Sarkissian CN, Scriver CR, Mamer OA (2000b) Measurement of phenyllactate, phenylacetate, and phenylpyruvate by negative ion chemical ionization-gas chromatography/mass spectrometry in brain of mouse genetic models of phenylketonuria and non-phenylketonuria hyperphenylalaninemia. Anal Biochem 280(2):242–249CrossRefPubMedGoogle Scholar
  27. Schindeler S, Ghosh-Jerath S, Thompson S et al (2007) The effects of large neutral amino acid supplements in PKU: an MRS and neuropsychological study. Mol Genet Metab 91(1):48–54CrossRefPubMedGoogle Scholar
  28. Scriver CR, Kaufman S (2001) Hyperphenylalaninemia: phenylalanine hydroxylase deficiency. In: Scriver CR, Beaudet AL, Sly WS, Valle D, (eds); Childs B, Kinzler KW, Vogelstein B (associated eds) The metabolic and molecular bases of inherited disease, 8th edn. McGraw-Hill, New York, pp 1667–1724Google Scholar
  29. Shefer S, Tint GS, Jean-Guillaume D et al (2000) Is there a relationship between 3-hydroxy-3-methylglutaryl coenzyme a reductase activity and forebrain pathology in the PKU mouse? J Neurosci Res 61(5):549–563CrossRefPubMedGoogle Scholar
  30. Silberberg DH (1967) Phenylketonuria metabolites in cerebellum culture morphology. Arch Neurol 17(5):524–529PubMedGoogle Scholar
  31. Smith CB, Kang J (2000) Cerebral protein synthesis in a genetic mouse model of phenylketonuria. Proc Natl Acad Sci U S A 97(20):11014–11019CrossRefPubMedGoogle Scholar
  32. Sommer I, Schachner M (1981) Monoclonal antibodies (O1 to O4) to oligodendrocyte cell surfaces: an immunocytological study in the central nervous system. Dev Biol 83(2):311–327CrossRefPubMedGoogle Scholar
  33. Stankoff B, Aigrot MS, Noel F, Wattilliaux A, Zalc B, Lubetzki C (2002) Ciliary neurotrophic factor (CNTF) enhances myelin formation: a novel role for CNTF and CNTF-related molecules. J Neurosci 22(21):9221–9227PubMedGoogle Scholar
  34. Surtees R, Blau N (2000) The neurochemistry of phenylketonuria. Eur J Pediatr 159(Suppl 2):S109–S113CrossRefPubMedGoogle Scholar
  35. van Spronsen FJ, Hoeksma M, Reijngoud DJ (2009) Brain dysfunction in phenylketonuria: is phenylalanine toxicity the only possible cause? J Inherit Metab Dis 32(1):46–51CrossRefPubMedGoogle Scholar
  36. Williams RA, Mamotte CD, Burnett JR (2008) Phenylketonuria: an inborn error of phenylalanine metabolism. Clin Biochem Rev 29(1):31–41PubMedGoogle Scholar
  37. Wu C, Schulte J, Sepp KJ, Littleton JT, Hong P (Submitted) Robust Neurite Detection and Automatic Morphological Analysis of Optical Microscopy Neuron Cell Culture Images in High-Content Screening. NeuroinformaticsGoogle Scholar
  38. Yamamura T, Konola JT, Wekerle H, Lees MB (1991) Monoclonal antibodies against myelin proteolipid protein: identification and characterization of two major determinants. J Neurochem 57(5):1671–1680CrossRefPubMedGoogle Scholar
  39. Zagreda L, Goodman J, Druin DP, McDonald D, Diamond A (1999) Cognitive deficits in a genetic mouse model of the most common biochemical cause of human mental retardation. J Neurosci 19(14):6175–6182PubMedGoogle Scholar

Copyright information

© SSIEM and Springer 2010

Authors and Affiliations

  • Renaud Schoemans
    • 1
  • Marie-Stéphane Aigrot
    • 2
  • Chaohong Wu
    • 3
  • Raphaël Marée
    • 4
  • Pengyu Hong
    • 3
  • Shibeshi Belachew
    • 5
  • Claire Josse
    • 1
  • Catherine Lubetzki
    • 2
  • Vincent Bours
    • 1
    • 6
    • 7
  1. 1.Human Genetics, GIGA-ResearchUniversity of LiègeLiègeBelgium
  2. 2.Cr-Icm, Inserm 711, UPMCParisFrance
  3. 3.Michtom School of Computer Science, Volen Center for Complex Systems, Room 261Brandeis UniversityWalthamUSA
  4. 4.Bioinformatics platform, GIGA-ResearchUniversity of LiègeLiègeBelgium
  5. 5.Neurosciences, GIGA-ResearchUniversity of LiègeLiègeBelgium
  6. 6.Genetics Center, CHU LiègeLiègeBelgium
  7. 7.Department of Genetics, CHU LiègeUniversité de Liège B34Liège, BelgiqueBelgium

Personalised recommendations