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Retinoic acid receptor β2 promotes functional regeneration of sensory axons in the spinal cord

Nature Neuroscience volume 9, pages 243250 (2006) | Download Citation

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Abstract

The embryonic CNS readily undergoes regeneration, unlike the adult CNS, which has limited axonal repair after injury. Here we tested the hypothesis that retinoic acid receptor β2 (RARβ2), critical in development for neuronal growth, may enable adult neurons to grow in an inhibitory environment. Overexpression of RARβ2 in adult rat dorsal root ganglion cultures increased intracellular levels of cyclic AMP and stimulated neurite outgrowth. Stable RARβ2 expression in DRG neurons in vitro and in vivo enabled their axons to regenerate across the inhibitory dorsal root entry zone and project into the gray matter of the spinal cord. The regenerated neurons enhanced second-order neuronal activity in the spinal cord, and RARβ2-treated rats showed highly significant improvement in sensorimotor tasks. These findings show that RARβ2 induces axonal regeneration programs within injured neurons and may thus offer new therapeutic opportunities for CNS regeneration.

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References

  1. 1.

    Retinoid signalling in the development of the central nervous system. Nat. Rev. Neurosci. 3, 843–853 (2002).

  2. 2.

    & Retinoids run rampant: multiple roles during spinal cord and motor neuron development. Neuron 40, 461–464 (2003).

  3. 3.

    Role and distribution of retinoic acid during CNS development. Int. Rev. Cytol. 209, 1–77 (2001).

  4. 4.

    , , & Activation of retinoic acid signalling after sciatic nerve injury: up-regulation of cellular retinoid binding proteins. Eur. J. Neurosci. 18, 1033–1040 (2003).

  5. 5.

    , , & The role of retinoic acid receptors in neurite outgrowth from different populations of embryonic mouse dorsal root ganglia. J. Cell Sci. 113, 2567–2574 (2000).

  6. 6.

    et al. Retinoic acid receptor β2 and neurite outgrowth in the adult mouse spinal cord in vitro. J. Cell Sci. 115, 3779–3786 (2002).

  7. 7.

    Nerve root damage and arachnoiditis. in Textbook of Pain (eds. Wall, P.D. & Melzack, R.) Ch. 38, 544–565 (Churchill Livingstone, Edinburgh, 1989).

  8. 8.

    Degeneration and Regeneration of the Nervous System (Hafner, New York, 1928).

  9. 9.

    Regenerating axons form nerve terminals at astrocytes. Brain Res. 347, 188–191 (1985).

  10. 10.

    et al. Rabies virus glycoprotein pseudotyping of lentiviral vectors enables retrograde axonal transport and access to the nervous system after peripheral delivery. Hum. Mol. Genet. 10, 2109–2121 (2001).

  11. 11.

    et al. Transduction patterns of pseudotyped lentiviral vectors in the nervous system. Mol. Ther. 9, 101–111 (2004).

  12. 12.

    , , , & Immunocytochemical localization of trkA receptors in chemically identified subgroups of adult rat sensory neurons. Eur. J. Neurosci. 7, 1484–1494 (1995).

  13. 13.

    et al. A distinct subgroup of small DRG cells express GDNF receptor components and GDNF is protective for these neurons after nerve injury. J. Neurosci. 18, 3059–3072 (1998).

  14. 14.

    , & Induction of c-fos-like protein in spinal cord neurons following sensory stimulation. Nature 328, 632–634 (1987).

  15. 15.

    , , & Nociceptive-specific activation of ERK in spinal neurons contributes to pain hypersensitivity. Nat. Neurosci. 2, 1114–1119 (1999).

  16. 16.

    , , & The “staircase test”: a measure of independent forelimb reaching and grasping abilities in rats. J. Neurosci. Methods 36, 219–228 (1991).

  17. 17.

    et al. The injured spinal cord spontaneously forms a new intraspinal circuit in adult rats. Nat. Neurosci. 7, 269–277 (2004).

  18. 18.

    et al. Neurite growth inhibitors restrict plasticity and functional recovery following corticospinal tract lesions. Nat. Neurosci. 1, 124–131 (1998).

  19. 19.

    , & Methods to assess the development and recovery of locomotor function after spinal cord injury in rats. Exp. Neurol. 119, 153–164 (1993).

  20. 20.

    & Repulsive factors and axon regeneration in the CNS. Curr. Opin. Neurobiol. 11, 89–94 (2001).

  21. 21.

    et al. An essential role for retinoid receptors RARbeta and RXRgamma in long-term potentiation and depression. Neuron 21, 1353–1361 (1998).

  22. 22.

    & Retinoic acid is detected at relatively high levels in the CNS of adult rats. Am. J. Physiol. Endocrinol. Metab. 282, E672–E678 (2002).

  23. 23.

    et al. Neuronal cyclic AMP controls the developmental loss in ability of axons to regenerate. J. Neurosci. 21, 4731–4739 (2001).

  24. 24.

    , , & Regeneration of sensory axons within the injured spinal cord induced by intraganglionic cAMP elevation. Neuron 34, 885–893 (2002).

  25. 25.

    et al. Spinal axon regeneration induced by elevation of cyclic AMP. Neuron 34, 895–903 (2002).

  26. 26.

    et al. Activated CREB is sufficient to overcome inhibitors in myelin and promote spinal axon regeneration in vivo. Neuron 44, 609–621 (2004).

  27. 27.

    A decade of molecular biology of retinoic acid receptors. FASEB J. 10, 940–954 (1996).

  28. 28.

    , & Transcriptional activation of Gs alpha expression by retinoic acid and parathyroid hormone-related protein in F9 teratocarcinoma cells. J. Biol. Chem. 265, 20081–20084 (1990).

  29. 29.

    , , & Different expression of adenylyl cyclase isoforms after retinoic acid induction of P19 teratocarcinoma cells. FEBS Lett. 415, 275–280 (1997).

  30. 30.

    & Cyclic AMP analogs and retinoic acid influence the expression of retinoic acid receptor alpha, beta, and gamma mRNAs in F9 teratocarcinoma cells. Mol. Cell. Biol. 10, 391–396 (1990).

  31. 31.

    , , & Functional regeneration of chronically injured sensory afferents into adult spinal cord after neurotrophin gene therapy. J. Neurosci. 21, 8408–8416 (2001).

  32. 32.

    , & Functional regeneration of sensory axons into the adult spinal cord. Nature 403, 312–316 (2000).

  33. 33.

    , , , & NT-3 delivered by an adenoviral vector induces injured dorsal root axons to regenerate into the spinal cord of adult rats. J. Neurosci. Res. 54, 554–562 (1998).

  34. 34.

    et al. Multicistronic lentiviral vector-mediated striatal gene transfer of aromatic L-amino acid decarboxylase, tyrosine hydroxylase, and GTP cyclohydrolase I induces sustained transgene expression, dopamine production, and functional improvement in a rat model of Parkinson's disease. J. Neurosci. 22, 10302–10312 (2002).

  35. 35.

    et al. Long-term replacement of a mutated nonfunctional CNS gene: reversal of hypothalamic diabetes insipidus using an EIAV-based lentiviral vector expressing arginine vasopressin. Mol. Ther. 7, 588–596 (2003).

  36. 36.

    et al. Stable gene transfer to the nervous system using a non-primate lentiviral vector. Gene Ther. 6, 1808–1818 (1999).

  37. 37.

    , , , & New methods to titrate EIAV-based lentiviral vectors. Mol. Ther. 5, 566–570 (2002).

  38. 38.

    , , & Growth responses of different subpopulations of adult sensory neurons to neurotrophic factors in vitro. Eur. J. Neurosci. 11, 3405–3414 (1999).

  39. 39.

    , , & Behaviour of DRG sensory neurites at the intact and injured adult rat dorsal root entry zone: postnatal neurites become paralysed, whilst injury improves the growth of embryonic neurites. Glia 26, 309–323 (1999).

  40. 40.

    , , , & Cloning of murine alpha and beta retinoic acid receptors and a novel receptor gamma predominantly expressed in skin. Nature 339, 714–717 (1989).

  41. 41.

    & Oligonucleotide probes for in situ hybridization. in In Situ Hybridization-A Practical Approach (ed. Wilkinson, D.G.) Ch. 2, 23–67 (Oxford Univ. Press, Oxford, 1998).

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Acknowledgements

We thank M. Agudo for help with the synthesis of the RARβ2 riboprobe, E. Foster for help with DRG cultures and L. Walmsley for technical help. This work was supported by Oxford BioMedica and the Medical Research Council.

Author information

Author notes

    • Liang-Fong Wong
    •  & Ping K Yip

    These authors contributed equally to this work.

Affiliations

  1. Oxford BioMedica (UK) Ltd., Medawar Centre, Robert Robinson Avenue, Oxford Science Park, Oxford OX4 4GA, UK.

    • Liang-Fong Wong
    • , Mimoun Azzouz
    • , Susan M Kingsman
    • , Alan J Kingsman
    •  & Nicholas D Mazarakis
  2. Neurorestoration Group, Wolfson Centre for Age-Related Diseases, Wolfson Wing, Hodgkin Building, King's College London, Guy's Campus, London SE1 1UL, UK.

    • Ping K Yip
    • , Anna Battaglia
    • , John Grist
    •  & Stephen B McMahon
  3. MRC Centre for Developmental Neurobiology, King's College London, New Hunts House, Guy's Campus, London SE1 1UL, UK.

    • Jonathan Corcoran
    •  & Malcolm Maden

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Competing interests

This Research has been funded by Oxford BioMedica. L.F.W., M.A., S.M.K., A.J.K and N.D.M. are employees of Oxford BioMedica.

Corresponding authors

Correspondence to Liang-Fong Wong or Nicholas D Mazarakis.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Morphology and inflammatory response in the rat spinal cord after vector injection.

  2. 2.

    Supplementary Fig. 2

    Increased cAMP immunoreactivity in EIAV-RARβ2 transduced DRG neurons.

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    Supplementary Fig. 3

    Comparison of functional recovery between EIAV-RARβ2 and cAMP-treated animals

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DOI

https://doi.org/10.1038/nn1622

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