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High cholesterol level is essential for myelin membrane growth

Abstract

Cholesterol in the mammalian brain is a risk factor for certain neurodegenerative diseases, raising the question of its normal function. In the mature brain, the highest cholesterol content is found in myelin. We therefore created mice that lack the ability to synthesize cholesterol in myelin-forming oligodendrocytes. Mutant oligodendrocytes survived, but CNS myelination was severely perturbed, and mutant mice showed ataxia and tremor. CNS myelination continued at a reduced rate for many months, and during this period, the cholesterol-deficient oligodendrocytes actively enriched cholesterol and assembled myelin with >70% of the cholesterol content of wild-type myelin. This shows that cholesterol is an indispensable component of myelin membranes and that cholesterol availability in oligodendrocytes is a rate-limiting factor for brain maturation.

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Figure 1: Inactivation of the squalene synthase gene (Fdft1) in oligodendrocytes.
Figure 2: Clinical phenotype.
Figure 3: Hypomyelination of CNS white matter tracks.
Figure 4: Ultrastructure of oligodendrocytes and myelin.
Figure 5: Biochemistry of mutant myelin.
Figure 6: Lipid analysis and detergent-resistant membranes.
Figure 7: Natural compensation for cholesterol deficiency.

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References

  1. Peters, A., Palay, S.L. & Webster, H.D. The Fine Structure of the Nervous System— the Neurons and Supporting Cells 3rd edn. (Oxford Univ. Press, New York, 1991).

    Google Scholar 

  2. Morell, P. & Jurevics, H. Origin of cholesterol in myelin. Neurochem. Res. 21, 463–470 (1996).

    Article  CAS  Google Scholar 

  3. Ohvo-Rekila, H., Ramstedt, B., Leppimaki, P. & Slotte, J.P. Cholesterol interactions with phospholipids in membranes. Prog. Lipid Res. 41, 66–97 (2002).

    Article  CAS  Google Scholar 

  4. Haines, T.H. Do sterols reduce proton and sodium leaks through lipid bilayers? Prog. Lipid Res. 40, 299–324 (2001).

    Article  CAS  Google Scholar 

  5. Brown, D.A. & London, E. Structure and function of sphingolipid- and cholesterol-rich membrane rafts. J. Biol. Chem. 275, 17221–17224 (2000).

    Article  CAS  Google Scholar 

  6. Simons, K. & Toomre, D. Lipid rafts and signal transduction. Nat. Rev. Mol. Cell Biol. 1, 31–39 (2000).

    Article  CAS  Google Scholar 

  7. Kramer, E.M., Schardt, A. & Nave, K.A. Membrane traffic in myelinating oligodendrocytes. Microsc. Res. Tech. 52, 656–671 (2001).

    Article  CAS  Google Scholar 

  8. Larocca, J.N. & Rodriguez-Gabin, A.G. Myelin biogenesis: vesicle transport in oligodendrocytes. Neurochem. Res. 27, 1313–1329 (2002).

    Article  CAS  Google Scholar 

  9. Simons, M., Kramer, E.M., Thiele, C., Stoffel, W. & Trotter, J. Assembly of myelin by association of proteolipid protein with cholesterol- and galactosylceramide-rich membrane domains. J. Cell Biol. 151, 143–154 (2000).

    Article  CAS  Google Scholar 

  10. Michikawa, M. & Yanagisawa, K. Inhibition of cholesterol production but not of nonsterol isoprenoid products induces neuronal cell death. J. Neurochem. 72, 2278–2285 (1999).

    Article  CAS  Google Scholar 

  11. Bradfute, D.L., Silva, C.J. & Simoni, R.D. Squalene synthase-deficient mutant of Chinese hamster ovary cells. J. Biol. Chem. 267, 18308–18314 (1992).

    CAS  PubMed  Google Scholar 

  12. Ohashi, K. et al. Early embryonic lethality caused by targeted disruption of the HMG-CoA reductase gene. J. Biol. Chem. 278, 42936–42941 (2003).

    Article  CAS  Google Scholar 

  13. Tozawa, R. et al. Embryonic lethality and defective neural tube closure in mice lacking squalene synthase. J. Biol. Chem. 274, 30843–30848 (1999).

    Article  CAS  Google Scholar 

  14. Gu, P., Ishii, Y., Spencer, T.A. & Shechter, I. Function-structure studies and identification of three enzyme domains involved in the catalytic activity in rat hepatic squalene synthase. J. Biol. Chem. 273, 12515–12525 (1998).

    Article  CAS  Google Scholar 

  15. Lappe-Siefke, C. et al. Disruption of Cnp1 uncouples oligodendroglial functions in axonal support and myelination. Nat. Genet. 33, 366–374 (2003).

    Article  CAS  Google Scholar 

  16. Genoud, S. et al. Notch1 control of oligodendrocyte differentiation in the spinal cord. J. Cell Biol. 158, 709–718 (2002).

    Article  CAS  Google Scholar 

  17. Belachew, S., Yuan, X. & Gallo, V. Unraveling oligodendrocyte origin and function by cell-specific transgenesis. Dev. Neurosci. 23, 287–298 (2001).

    Article  CAS  Google Scholar 

  18. Gallyas, F. Silver staining of myelin by means of physical development. Neurol. Res. 1, 203–209 (1979).

    Article  CAS  Google Scholar 

  19. Salzer, J.L. Polarized domains of myelinated axons. Neuron 40, 297–318 (2003).

    Article  CAS  Google Scholar 

  20. Marcus, J. & Popko, B. Galactolipids are molecular determinants of myelin development and axo-glial organization. Biochim. Biophys. Acta 1573, 406–413 (2002).

    Article  CAS  Google Scholar 

  21. Stoffel, W. & Bosio, A. Myelin glycolipids and their functions. Curr. Opin. Neurobiol. 7, 654–661 (1997).

    Article  CAS  Google Scholar 

  22. Nave, K.A. & Griffiths, I.R. Models of Pelizaeus-Merzbacher disease. in Myelin Biology and Disorders Vol. 2 (ed. Lazzarini, R.A.) 1125–1142 (Academic, London, 2003).

    Google Scholar 

  23. Sandhoff, R., Brügger, B., Jeckel, D., Lehmann, W.D. & Wieland, F.T. Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry. J. Lipid Res. 40, 126–132 (1999).

    CAS  PubMed  Google Scholar 

  24. Simons, M. et al. Overexpression of the myelin proteolipid protein leads to accumulation of cholesterol and proteolipid protein in endosomes/lysosomes: implications for Pelizaeus-Merzbacher disease. J. Cell Biol. 157, 327–336 (2002).

    Article  CAS  Google Scholar 

  25. Readhead, C. et al. Expression of a myelin basic protein gene in transgenic shiverer mice: correction of the dysmyelinating phenotype. Cell 48, 703–712 (1987).

    Article  CAS  Google Scholar 

  26. Pfrieger, F.W. Outsourcing in the brain: do neurons depend on cholesterol delivery by astrocytes? Bioessays 25, 72–78 (2003).

    Article  Google Scholar 

  27. Herz, J. & Bock, H.H. Lipoprotein receptors in the nervous system. Annu. Rev. Biochem. 71, 405–434 (2002).

    Article  CAS  Google Scholar 

  28. Lu, Q.R. et al. Sonic hedgehog–regulated oligodendrocyte lineage genes encoding bHLH proteins in the mammalian central nervous system. Neuron 25, 317–329 (2000).

    Article  CAS  Google Scholar 

  29. Alberta, J.A. et al. Sonic hedgehog is required during an early phase of oligodendrocyte development in mammalian brain. Mol. Cell. Neurosci. 18, 434–441 (2001).

    Article  CAS  Google Scholar 

  30. Cooper, M.K. et al. A defective response to Hedgehog signaling in disorders of cholesterol biosynthesis. Nat. Genet. 33, 508–513 (2003).

    Article  CAS  Google Scholar 

  31. Nwokoro, N.A., Wassif, C.A. & Porter, F.D. Genetic disorders of cholesterol biosynthesis in mice and humans. Mol. Genet. Metab. 74, 105–119 (2001).

    Article  CAS  Google Scholar 

  32. Kim, S.U. Effects of the cholesterol biosynthesis inhibitor ay9944 on organotypic cultures ofmouse spinal cord. Retarded myelinogenesis and induction of cytoplasmic inclusions. Lab. Invest. 32, 720–728 (1975).

    CAS  PubMed  Google Scholar 

  33. Brown, M.S. & Goldstein, J.L. A proteolytic pathway that controls the cholesterol content of membranes, cells, and blood. Proc. Natl. Acad. Sci. USA 96, 11041–11048 (1999).

    Article  CAS  Google Scholar 

  34. Verheijen, M.H., Chrast, R., Burrola, P. & Lemke, G. Local regulation of fat metabolism in peripheral nerves. Genes Dev. 17, 2450–2464 (2003).

    Article  CAS  Google Scholar 

  35. Sereda, M.W., Meyer Zu, H.G., Suter, U., Uzma, N. & Nave, K.A. Therapeutic administration of progesterone antagonist in a model of Charcot-Marie-Tooth disease (CMT-1A). Nat. Med. 9, 1533–1537 (2003).

    Article  CAS  Google Scholar 

  36. Lazzarini, R.A. Myelin Biology and Disorders (Academic, London, 2003).

    Google Scholar 

  37. Popko, B. et al. Myelin deficient mice: expression of myelin basic protein and generation of mice with varying levels of myelin. Cell 48, 713–721 (1987).

    Article  CAS  Google Scholar 

  38. Bjorkhem, I. & Meaney, S. Brain cholesterol: long secret life behind a barrier. Arterioscler. Thromb. Vasc. Biol. 24, 806–815 (2004).

    Article  Google Scholar 

  39. Wechsler, A. et al. Generation of viable cholesterol-free mice. Science 302, 2087 (2003).

    Article  CAS  Google Scholar 

  40. Joyner, A.L. Gene targeting and gene trap screens using embryonic stem cells: new approaches to mammalian development. Bioessays 13, 649–656 (1991).

    Article  CAS  Google Scholar 

  41. Jones, B.J. & Roberts, D.J. A rotarod suitable for quantitative measurements of motor incoordination in naive mice. Naunyn Schmiedebergs Arch. Exp. Pathol. Pharmakol. 259, 211 (1968).

    Article  CAS  Google Scholar 

  42. Schaeren-Wiemers, N. & Gerfin-Moser, A. A single protocol to detect transcripts of various types and expression levels in neural tissue and cultured cells: in situ hybridization using digoxigenin-labelled cRNA probes. Histochemistry 100, 431–440 (1993).

    Article  CAS  Google Scholar 

  43. Huber, A.B., Weinmann, O., Brosamle, C., Oertle, T. & Schwab, M.E. Patterns of Nogo mRNA and protein expression in the developing and adult rat and after CNS lesions. J. Neurosci. 22, 3553–3567 (2002).

    Article  CAS  Google Scholar 

  44. Jung, M., Sommer, I., Schachner, M. & Nave, K.A. Monoclonal antibody O10 defines a conformationally sensitive cell-surface epitope of proteolipid protein (PLP): evidence that PLP misfolding underlies dysmyelination in mutant mice. J. Neurosci. 16, 7920–7929 (1996).

    Article  CAS  Google Scholar 

  45. Norton, W.T. & Poduslo, S.E. Myelination in rat brain: method of myelin isolation. J. Neurochem. 21, 749–757 (1973).

    Article  CAS  Google Scholar 

  46. Brown, D.A. & Rose, J.K. Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell 68, 533–544 (1992).

    Article  CAS  Google Scholar 

  47. Jurevics, H. et al. Normal metabolism but different physical properties of myelin from mice deficient in proteolipid protein. J. Neurosci. Res. 71, 826–834 (2003).

    Article  CAS  Google Scholar 

  48. Plonne, D. et al. Separation of the intracellular secretory compartment of rat liver and isolated rat hepatocytes in a single step using self-generating gradients of iodixanol. Anal. Biochem. 276, 88–96 (1999).

    Article  CAS  Google Scholar 

  49. Bligh, E.G. & Dyer, W.J. A rapid method of total lipid extraction and purification. Can. J. Med. Sci. 37, 911–917 (1959).

    CAS  Google Scholar 

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Acknowledgements

We dedicate this work to P. Morell for his pioneering work on brain cholesterol synthesis. We thank L. Bitterberg, A. Fahrenholz, S. Keese, I. Leibrecht, E. Nicksch and M. Schindler for excellent technical help; M. Schwab for providing us with antibodies; and M. Simons for helpful comments on the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft (SFB523 to K.A.N.; Wi654/7 to B.B. and F.W.), the Hertie Institute of Multiple Sclerosis Research and grants from the European Union (K.A.N.).

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Correspondence to Klaus-Armin Nave.

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Supplementary information

Supplementary Fig. 1

Loss of SQS in mutant primary oligodendrocytes. (PDF 51 kb)

Supplementary Fig. 2

Measurement of cholesterol in myelin. (PDF 26 kb)

Supplementary Fig. 3

Reduction of myelin mRNAs during postnatal development. (PDF 10 kb)

Supplementary Fig. 4

Astroglial cells as candidates to provide cholesterol. (PDF 73 kb)

Supplementary Fig. 5

No 'rescue' by cholesterol-rich diet. (PDF 54 kb)

Supplementary Table 1

Lipid content in purified myelin. (PDF 9 kb)

Supplementary Video 1

Mutants at 20 days of age show strong tremors and ataxia. (MOV 1104 kb)

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Saher, G., Brügger, B., Lappe-Siefke, C. et al. High cholesterol level is essential for myelin membrane growth. Nat Neurosci 8, 468–475 (2005). https://doi.org/10.1038/nn1426

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