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Intra-axonal translation and retrograde trafficking of CREB promotes neuronal survival

Nature Cell Biology volume 10, pages 149159 (2008) | Download Citation

Abstract

During development of the nervous system, axons and growth cones contain mRNAs such as β-actin, cofilin and RhoA, which are locally translated in response to guidance cues. Intra-axonal translation of these mRNAs results in local morphological responses; however, other functions of intra-axonal mRNA translation remain unknown. Here, we show that axons of developing mammalian neurons contain mRNA encoding the cAMP-responsive element (CRE)-binding protein (CREB). CREB is translated within axons in response to nerve growth factor (NGF) and is retrogradely trafficked to the cell body. In neurons that are selectively deficient in axonal CREB transcripts, increases in nuclear pCREB, CRE-mediated transcription and neuronal survival elicited by axonal application of NGF are abolished, indicating a signalling function for axonally synthesized CREB. These studies identify a signalling role for axonally derived CREB, and indicate that signal-dependent synthesis and retrograde trafficking of transcription factors enables specific transcriptional responses to signalling events at distal axons.

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References

  1. 1.

    & Pathways for mRNA localization in the cytoplasm. Trends Biochem. Sci. 31, 687–693 (2006).

  2. 2.

    & RNA translation in axons. Annu Rev. Cell Dev. Biol. 20, 505–523 (2004).

  3. 3.

    et al. Sorting of ß-actin mRNA and protein to neurites and growth cones in culture. J. Neurosci. 18, 251–265 (1998).

  4. 4.

    et al. Local translation of RhoA regulates growth cone collapse. Nature 436, 1020–1024 (2005).

  5. 5.

    et al. Signaling mechanisms underlying Slit2-induced collapse of Xenopus retinal growth cones. Neuron 49, 215–228 (2006).

  6. 6.

    et al. GDNF acts through PEA3 to regulate cell body positioning and muscle innervation of specific motor neuron pools. Neuron 35, 893–905 (2002).

  7. 7.

    , & Functions and mechanisms of retrograde neurotrophin signalling. Nature Rev. Neurosci. 6, 615–625 (2005).

  8. 8.

    et al. Peripheral NT3 signaling is required for ETS protein expression and central patterning of proprioceptive sensory afferents. Neuron 38, 403–416 (2003).

  9. 9.

    et al. Retrograde BMP signaling regulates trigeminal sensory neuron identities and the formation of precise face maps. Neuron 55, 572–586 (2007).

  10. 10.

    & Long-distance retrograde neurotrophic signaling. Curr. Opin. Neurobiol. 15, 40–48 (2005).

  11. 11.

    & Regulation of cap-dependent translation by eIF4E inhibitory proteins. Nature 433, 477–480 (2005).

  12. 12.

    Local control of neurite development by nerve growth factor. Proc. Natl Acad. Sci. USA 74, 4516–4519 (1977).

  13. 13.

    , , & Apoptosis, axonal growth defects, and degeneration of peripheral neurons in mice lacking CREB. Neuron 34, 371–385 (2002).

  14. 14.

    , , & Radiation-induced apoptosis in dorsal root ganglion neurons. J. Neurocytol. 26, 771–777 (1997).

  15. 15.

    et al. Inhibitors of protein synthesis and RNA synthesis prevent neuronal death caused by nerve growth factor deprivation. J. Cell Biol. 106, 829–844 (1988).

  16. 16.

    , , & Chromatin cleavage in apoptosis: association with condensed chromatin morphology and dependence on macromolecular synthesis. J. Pathol. 142, 67–77 (1984).

  17. 17.

    et al. A functional role for intra-axonal protein synthesis during axonal regeneration from adult sensory neurons. J. Neurosci. 21, 9291–9303 (2001).

  18. 18.

    et al. Single-cell transcriptional analysis of neuronal progenitors. Neuron 38, 161–175 (2003).

  19. 19.

    , , , & Targeting of the CREB gene leads to up-regulation of a novel CREB mRNA isoform. EMBO J. 15, 1098–1106 (1996).

  20. 20.

    & Localization and active transport of mRNA in axons of sympathetic neurons in culture. J. Neurosci. 16, 1346–1358 (1996).

  21. 21.

    et al. Presence and phosphorylation of transcription factors in developing dendrites. Proc. Natl Acad. Sci. USA 95, 2313–2318 (1998).

  22. 22.

    , , , & Dynamic visualization of local protein synthesis in hippocampal neurons. Neuron 30, 489–502 (2001).

  23. 23.

    , , , & A modified Sindbis vector for prolonged gene expression in neurons. J. Neurophysiol. 90, 2741–2745 (2003).

  24. 24.

    , , , & Importin α: a multipurpose nuclear-transport receptor. Trends Cell Biol. 14, 505–514 (2004).

  25. 25.

    et al. Axoplasmic importins enable retrograde injury signaling in lesioned nerve. Neuron 40, 1095–1104 (2003).

  26. 26.

    & Nuclear translocation and DNA recognition signals colocalized within the bZIP domain of cyclic adenosine 3′,5′-monophosphate response element-binding protein CREB. Mol. Endocrinol. 5, 1418–1430 (1991).

  27. 27.

    et al. Monomeric green-to-red photoconvertible fluorescent protein activated by a visible blue light. Nature Biotechnol. 24, 461–465 (2006).

  28. 28.

    , & Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype. J. Cell Biol. 127, 441–451 (1994).

  29. 29.

    et al. Neurotrophin-induced transport of a β-actin mRNP complex increases β-actin levels and stimulates growth cone motility. Neuron 31, 261–275 (2001).

  30. 30.

    & Rapid retrograde transport of dopamine-β-hydroxylase as examined by the stop-flow technique. Brain Res. 102, 217–228 (1976).

  31. 31.

    & Retrograde transport and steady-state distribution of 125I-nerve growth factor in rat sympathetic neurons in compartmented cultures. J. Neurosci. 17, 1282–1290 (1997).

  32. 32.

    , , & The effect of acrylamide and other sulfhydryl alkylators on the ability of dynein and kinesin to translocate microtubules in vitro. Toxicol. Appl. Pharmacol. 133, 73–81 (1995).

  33. 33.

    et al. Neurotrophins use the Erk5 pathway to mediate a retrograde survival response. Nature Neurosci. 4, 981–988 (2001).

  34. 34.

    et al. One at a time, live tracking of NGF axonal transport using quantum dots. Proc. Natl Acad. Sci. USA 104, 13666–13671 (2007).

  35. 35.

    , , & Functional and selective RNA interference machinery in axonal growth cones. J. Neurosci. 26, 5727–5732 (2006).

  36. 36.

    et al. A neurotrophin signaling cascade coordinates sympathetic neuron development through differential control of TrkA trafficking and retrograde signaling. Cell 118, 243–255 (2004).

  37. 37.

    , , & An NGF–TrkA-mediated retrograde signal to transcription factor CREB in sympathetic neurons. Science 277, 1097–1100 (1997).

  38. 38.

    et al. Rapid nuclear responses to target-derived neurotrophins require retrograde transport of ligand-receptor complex. J. Neurosci. 19, 7889–7900 (1999).

  39. 39.

    , & Compartmentalization of growth factor receptor signalling. Curr. Opin. Cell Biol. 17, 107–111 (2005).

  40. 40.

    , , & NGF signaling from clathrin-coated vesicles: evidence that signaling endosomes serve as a platform for the Ras–MAPK pathway. Neuron 32, 801–814 (2001).

  41. 41.

    et al. NGF signaling in sensory neurons: evidence that early endosomes carry NGF retrograde signals. Neuron 39, 69–84 (2003).

  42. 42.

    et al. CREB transcriptional activity in neurons is regulated by multiple, calcium-specific phosphorylation events. Neuron 34, 221–233 (2002).

  43. 43.

    , & What turns CREB on? Cell Signal 16, 1211–1227 (2004).

  44. 44.

    , , , & NF-κB functions in synaptic signaling and behavior. Nature Neurosci. 6, 1072–1078 (2003).

  45. 45.

    & Nervy links protein kinase a to plexin-mediated semaphorin repulsion. Science 303, 1204–1207 (2004).

  46. 46.

    , , & Specification of neuropeptide cell identity by the integration of retrograde BMP signaling and a combinatorial transcription factor code. Cell 113, 73–86 (2003).

  47. 47.

    el al. Retrograde Gbb signaling through the Bmp type 2 receptor wishful thinking regulates systemic FMRFa expression in Drosophila. Development 130, 5457–5470 (2003).

  48. 48.

    , , & Functional inhibition of the p75 receptor using a small interfering RNA. Biochem. Biophys. Res. Comm. 301, 804–809 (2003).

  49. 49.

    & Neuronal activity regulates diffusion across the neck of dendritic spines. Science 310, 866–869 (2005).

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Acknowledgements

We thank: D. Fischman for suggesting the use of Boyden chambers; K. Wu for advice and assistance; S. Schlesinger and A. Jeromin for Sindbis plasmids; R. Ratan for CRE plasmids; and A. North for advice and assistance with Dendra photoactivation. Supported by the Paralyzed Veterans of America (L.J.C), the National Institute of Mental Health, the National Alliance for Autism Research, the Klingenstein Foundation, and a Charles A. Dana foundation grant to Weill-Cornell (S.R.J), and the European Commission FP-6 Integrated Project LSHG-CT-2003-503259 (K.A.L).

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Affiliations

  1. Department of Pharmacology, Weill Medical College, Cornell University, New York, NY 10021, USA.

    • Llewellyn J. Cox
    • , Ulrich Hengst
    •  & Samie R. Jaffrey
  2. Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, Moscow 117997, Russia.

    • Nadya G. Gurskaya
    •  & Konstantin A. Lukyanov

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Contributions

L.J.C and S.R.J. designed the experiments, analysed the data and wrote the manuscript. L.J.C. carried out the experiments, collected data and prepared the figures. N.G.G. and K.A.L. provided reagents and input for the experimental design of photoactivation experiments. U.H. designed, carried out and analysed experiments using the translational reporter and prepared figures. S.R.J. directed and planned the project.

Note: An error in the original AOP version of the article has now been corrected in both the HTML and PDF versions; the article will appear correctly in the February print issue of Nature Cell Biology.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Samie R. Jaffrey.

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    Supplementary Figures S1, S2 and S3, Supplementary Tables S1, S2, S3 and S4 and Supplementary Methods

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DOI

https://doi.org/10.1038/ncb1677

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