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