Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Cyclic AMP controls BDNF-induced TrkB phosphorylation and dendritic spine formation in mature hippocampal neurons

Abstract

Synaptic actions of brain-derived neurotrophic factor (BDNF) are 'gated' by cyclic AMP (cAMP), but the underlying molecular mechanisms remain unclear. Here we report that cAMP regulates BDNF function in mature hippocampal neurons by modulating the signaling and trafficking of its receptor TrkB. cAMP gated the TrkB tyrosine kinase with three characteristic features: BDNF-induced TrkB phosphorylation was attenuated by inhibitors of cAMP signaling, it was potentiated by cAMP analogs, and activation of the cAMP pathway alone had no effect. In addition, cAMP facilitated trafficking of TrkB to dendritic spines, possibly by promoting its interaction with synaptic scaffolding protein PSD-95. Norepinephrinergic and dopaminergic agonists, which elevate intracellular cAMP concentration, also enhanced TrkB phosphorylation and its translocation to spines. cAMP gated long-term modulation by BDNF of spine density, but not the number of primary dendrites. These results reveal a specific role of cAMP in controlling BDNF actions in the brain, and provide new insights into the molecular mechanism underlying cAMP gating.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Purchase on Springer Link

Instant access to full article PDF

Prices may be subject to local taxes which are calculated during checkout

Figure 1: cAMP facilitates the increase in dendritic spine density induced by BDNF.
Figure 2: cAMP is not involved in the BDNF regulation of dendritic growth.
Figure 3: Role of cAMP in BDNF modulation of HFS-induced synaptic fatigue and LTP at hippocampal CA1 synapses.
Figure 4: Regulation of TrkB phosphorylation at Tyr 490 by cAMP in mature hippocampal neurons.
Figure 5: Specificity of cAMP regulation of TrkB phosphorylation in mature hippocampal neurons.
Figure 6: cAMP signaling does not alter cell surface expression of TrkB.
Figure 7: cAMP facilitates the colocalization of TrkB with PSD-95.
Figure 8: cAMP enhances the association of TrkB with PSD-95.

Similar content being viewed by others

References

  1. Lu, B. Acute and long-term regulation of synapses by neurotrophins. Prog. Brain Res. 146, 137–150 (2004).

    CAS  PubMed  Google Scholar 

  2. Poo, M.M. Neurotrophins as synaptic modulators. Nat. Rev. Neurosci. 2, 24–32 (2001).

    Article  CAS  Google Scholar 

  3. Lessmann, V., Gottmann, K. & Heumann, R. BDNF and NT-4/5 enhance glutamatergic synaptic transmission in cultured hippocampal neurons. Neuroreport 6, 21–25 (1994).

    Article  CAS  Google Scholar 

  4. Takei, N. et al. Brain-derived neurotrophic factor increases the stimulation-evoked release of glutamate and the levels of exocytosis-associated proteins in cultured cortical neurons from embryonic rats. J. Neurochem. 68, 370–375 (1997).

    Article  CAS  Google Scholar 

  5. Figurov, A., Pozzo-Miller, L., Olafsson, P., Wang, T. & Lu, B. Regulation of synaptic responses to high-frequency stimulation and LTP by neurotrophins in the hippocampus. Nature 381, 706–709 (1996).

    Article  CAS  Google Scholar 

  6. Cohen-Cory, S. & Fraser, S.E. Effects of brain-derived neurotrophic factor on optic axon branching and remodelling in vivo. Nature 378, 192–196 (1995).

    Article  CAS  Google Scholar 

  7. Gallo, G. & Letourneau, P.C. Localized sources of neurotrophins initiate axon collateral sprouting. J. Neurosci. 18, 5403–5414 (1998).

    Article  CAS  Google Scholar 

  8. McAllister, A.M., Katz, L.C. & Lo, D.C. Neurotrophins and synaptic plasticity. Annu. Rev. Neurosci. 22, 295–318 (1999).

    Article  CAS  Google Scholar 

  9. Cabelli, R.J., Horn, A. & Shatz, C.J. Inhibition of ocular dominance column formation by infusion of NT-4/5 or BDNF. Science 267, 1662–1666 (1995).

    Article  CAS  Google Scholar 

  10. Tyler, W.J. & Pozzo-Miller, L.D. BDNF enhances quantal neurotransmitter release and increases the number of docked vesicles at the active zones of hippocampal excitatory synapses. J. Neurosci. 21, 4249–4258 (2001).

    Article  CAS  Google Scholar 

  11. Shimada, A., Mason, C.A. & Morrison, M.E. TrkB signaling modulates spine density and morphology independent of dendrite structure in cultured neonatal Purkinje cells. J. Neurosci. 18, 8559–8570 (1998).

    Article  CAS  Google Scholar 

  12. Huang, E.J. & Reichardt, L.F. Trk receptors: roles in neuronal signal transduction. Annu. Rev. Biochem. 72, 609–642 (2003).

    Article  CAS  Google Scholar 

  13. Meyer-Franke, A., Kaplan, M.R., Pfrieger, F.W. & Barres, B.A. Characterization of the signaling interactions that promote the survival and growth of developing retinal ganglion cells in culture. Neuron 15, 805–819 (1995).

    Article  CAS  Google Scholar 

  14. Song, H.J., Ming, G.L. & Poo, M.M. cAMP-induced switching in turning direction of nerve growth cones. Nature 388, 275–279 (1997).

    Article  CAS  Google Scholar 

  15. Gaiddon, C., Loeffler, J.P. & Larmet, Y. Brain-derived neurotrophic factor stimulates AP-1 and cyclic AMP-responsive element dependent transcriptional activity in central nervous system neurons. J. Neurochem. 66, 2279–2286 (1996).

    Article  CAS  Google Scholar 

  16. Brewer, G.J. Serum-free B27/neurobasal medium supports differentiated growth of neurons from the striatum, substantia nigra, septum, cerebral cortex, cerebellum, and dentate gyrus. J. Neurosci. Res. 42, 674–683 (1995).

    Article  CAS  Google Scholar 

  17. Tartaglia, N. et al. Protein synthesis-dependent and -independent regulation of hippocampal synapses by brain-derived neurotrophic factor. J. Biol. Chem. 276, 37585–37593 (2001).

    Article  CAS  Google Scholar 

  18. Boulanger, L. & Poo, M. Gating of BDNF-induced synaptic potentiation by cAMP. Science 284, 1982–1984 (1999).

    Article  CAS  Google Scholar 

  19. Wu, K. et al. Functional trkB neurotrophin receptors are intrinsic components of the adult brain postsynaptic density. Brain Res. Mol. Brain Res. 43, 286–290 (1996).

    Article  CAS  Google Scholar 

  20. Harris, K.M., Jensen, F.E. & Tsao, B. Three-dimensional structure of dendritic spines and synapses in rat hippocampus (CA1) at postnatal day 15 and adult ages: implications for the maturation of synaptic physiology and long-term potentiation. J. Neurosci. 12, 2685–2705 (1992).

    Article  CAS  Google Scholar 

  21. Ziv, N.E. & Smith, S.J. Evidence for a role of dendritic filopodia in synaptogenesis and spine formation. Neuron 17, 91–102 (1996).

    Article  CAS  Google Scholar 

  22. Papa, M., Bundman, M.C., Greenberger, V. & Segal, M. Morphological analysis of dendritic spine development in primary cultures of hippocampal neurons. J. Neurosci. 15, 1–11 (1995).

    Article  CAS  Google Scholar 

  23. Iyengar, R. Gating by cyclic AMP: expanded role for an old signaling pathway. Science 271, 461–463 (1996).

    Article  CAS  Google Scholar 

  24. McAllister, A.K., Lo, D.C. & Katz, L.C. Neurotrophins regulate dendritic growth in developing visual cortex. Neuron 15, 791–803 (1995).

    Article  CAS  Google Scholar 

  25. Gottschalk, W., Pozzo-Miller, L.D., Figurov, A. & Lu, B. Presynaptic modulation of synaptic transmission and plasticity by brain-derived neurotrophic factor in the developing hippocampus. J. Neurosci. 18, 6830–6839 (1998).

    Article  CAS  Google Scholar 

  26. Blitzer, R.D., Wong, T., Nouranifar, R., Iyengar, R. & Landau, E.M. Postsynaptic cAMP pathway gates early LTP in hippocampal CA1 region. Neuron 15, 1403–1414 (1995).

    Article  CAS  Google Scholar 

  27. Lezcano, N. & Bergson, C. D1/D5 dopamine receptors stimulate intracellular calcium release in primary cultures of neocortical and hippocampal neurons. J. Neurophysiol. 87, 2167–2175 (2002).

    Article  CAS  Google Scholar 

  28. Brinton, R.D., Thompson, R.H. & Brownson, E.A. Spatial, cellular and temporal basis of vasopressin potentiation of norepinephrine-induced cAMP formation. Eur. J. Pharmacol. 405, 73–88 (2000).

    Article  CAS  Google Scholar 

  29. Meyer-Franke, A. et al. Depolarization and cAMP elevation rapidly recruit TrkB to the plasma membrane of CNS neurons. Neuron 21, 681–693 (1998).

    Article  CAS  Google Scholar 

  30. Du, J., Feng, L., Yang, F. & Lu, B. Activity- and Ca2+-dependent modulation of surface expression of brain-derived neurotrophic factor receptors in hippocampal neurons. J. Cell Biol. 150, 1423–1434 (2000).

    Article  CAS  Google Scholar 

  31. Haapasalo, A. et al. Regulation of TRKB surface expression by brain-derived neurotrophic factor and truncated TRKB isoforms. J. Biol. Chem. 277, 43160–43167 (2002).

    Article  CAS  Google Scholar 

  32. Kryl, D. et al. Subcellular localization of full-length and truncated Trk receptor isoforms in polarized neurons and epithelial cells. J. Neurosci. 19, 5823–5833 (1999).

    Article  CAS  Google Scholar 

  33. Hu, B.R. et al. Assembly of proteins to postsynaptic densities after transient cerebral ischemia. J. Neurosci. 18, 625–633 (1998).

    Article  CAS  Google Scholar 

  34. Kornau, H.C., Schenker, L.T., Kennedy, M.B. & Seeburg, P.H. Domain interaction between NMDA receptor subunits and the postsynaptic density protein PSD-95. Science 269, 1737–1740 (1995).

    Article  CAS  Google Scholar 

  35. Chen, J. & Iyengar, R. Suppression of Ras-induced transformation of NIH 3T3 cells by activated G alpha s. Science 263, 1278–1281 (1994).

    Article  CAS  Google Scholar 

  36. Blitzer, R.D. et al. Gating of CaMKII by cAMP-regulated protein phosphatase activity during LTP. Science 280, 1940–1942 (1998).

    Article  CAS  Google Scholar 

  37. Tartaglia, N. et al. Protein synthesis dependent and independent regulation of hippocampal synapses by brain-derived neurotrophic factor. J. Biol. Chem. 276, 37585–37593 (2001).

    Article  CAS  Google Scholar 

  38. Drake, C.T., Milner, T.A. & Patterson, S.L. Ultrastructural localization of full-length trkB immunoreactivity in rat hippocampus suggests multiple roles in modulating activity-dependent synaptic plasticity. J. Neurosci. 19, 8009–8026 (1999).

    Article  CAS  Google Scholar 

  39. Patterson, S.L. et al. Some forms of cAMP-mediated long-lasting potentiation are associated with release of BDNF and nuclear translocation of phospho-MAP kinase. Neuron 32, 123–140 (2001).

    Article  CAS  Google Scholar 

  40. Wyneken, U. et al. Kainate-induced seizures alter protein composition and N-methyl-D-aspartate receptor function of rat forebrain postsynaptic densities. Neuroscience 102, 65–74 (2001).

    Article  CAS  Google Scholar 

  41. Scannevin, R.H. & Huganir, R.L. Postsynaptic organization and regulation of excitatory synapses. Nat. Rev. Neurosci. 1, 133–141 (2000).

    Article  CAS  Google Scholar 

  42. Sheng, M. Molecular organization of the postsynaptic specialization. Proc. Natl. Acad. Sci. USA 98, 7058–7061 (2001).

    Article  CAS  Google Scholar 

  43. O'Brien, R.J., Lau, L.F. & Huganir, R.L. Molecular mechanisms of glutamate receptor clustering at excitatory synapses. Curr. Opin. Neurobiol. 8, 364–369 (1998).

    Article  CAS  Google Scholar 

  44. Sala, C. et al. Regulation of dendritic spine morphology and synaptic function by Shank and Homer. Neuron 31, 115–130 (2001).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank Regeneron Pharmaceuticals for providing recombinant BDNF. This work was supported by funds from the National Institute of Child Heath and Human Development (NICHD) intramural program, Major State Basic Research Program of China (No. G2000077800) and National Natural Science Foundation of China (No. 30228020 and No. 30470533).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Bai Lu.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Forskolin facilitates the increase in dendritic protrusion density induced by BDNF. (GIF 23 kb)

Supplementary Fig. 2

Regulation of TrkB phosphorylation at Tyr 490 by forskolin. (GIF 42 kb)

Supplementary Fig. 3

The level of endogenous BDNF does not affect TrkB phosphorylation and the increase of spine density induced by BDNF. (GIF 32 kb)

Supplementary Video 1

Spine dynamics under control conditions. Cultured hippocampal neurons were transfected with GFP at day 7 and observed at day 21 in vitro. Neurons were placed in a chamber on the heated stage of a Zeiss confocal microscope (40, NA 1.30, 488 nm laser), incubated in L-15 medium containing B27 supplement. Images were captured at the indicated time for up to 15 hours to generate 13-15 video frames. Under control conditions most spines were relatively stable. There was only a bit of head morphing, but no protrusive motility at all. (MOV 1395 kb)

Supplementary Video 2

Spine dynamics in the presence of 5 ng/ml BDNF. Appearance of the yellow dot denotes addition of BDNF. Spine density was increased compared to the control condition. New spines emerged, disappeared and reemerged in the same location, and protrusion motility of new spines was observed (arrows). (MOV 669 kb)

Supplementary Video 3

Spine dynamics in the presence of 10 M Sp-cAMP. Appearance of the red dot denotes addition of Sp-cAMP. Spine motility was significantly increased compared to the control condition. Spines disappeared and then reemerged in new or previous locations. (MOV 517 kb)

Supplementary Video 4

Spine dynamics in the presence of 5 ng/ml BDNF and 10 M Sp-cAMP. The red dot denotes addition of Sp-cAMP, the yellow dot denotes addition of BDNF. Compared to BDNF alone (supplementary video 2), more spines appeared and most new spines exhibited rapid head morphing (arrows). New spines were maintained in the same location over 24 hours. (MOV 649 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ji, Y., Pang, P., Feng, L. et al. Cyclic AMP controls BDNF-induced TrkB phosphorylation and dendritic spine formation in mature hippocampal neurons. Nat Neurosci 8, 164–172 (2005). https://doi.org/10.1038/nn1381

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nn1381

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing