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AKAP-mediated feedback control of cAMP gradients in developing hippocampal neurons

Nature Chemical Biology volume 13, pages 425–431 (2017)Cite this article

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Abstract

Cyclic AMP (cAMP) and protein kinase A (PKA), classical examples of spatially compartmentalized signaling molecules, are critical axon determinants that regulate neuronal polarity and axon formation, yet little is known about micro-compartmentalization of cAMP and PKA signaling and its role in developing neurons. Here, we revealed that cAMP forms a gradient in developing hippocampal neurons, with higher cAMP levels in more distal regions of the axon compared to other regions of the cell. Interestingly, this cAMP gradient changed according to the developmental stage and depended on proper anchoring of PKA by A-kinase anchoring proteins (AKAPs). Disrupting PKA anchoring to AKAPs increased the cAMP gradient in early-stage neurons and led to enhanced axon elongation. Our results provide new evidence for a local negative-feedback loop, assembled by AKAPs, for the precise control of a growth-stage-dependent cAMP gradient to ensure proper axon growth.

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Figure 1: Spatial regulation of cAMP production in early polarized hippocampal neurons.
Figure 2: Spatial regulation of PKA activity in early polarized hippocampal neurons.
Figure 3: Acute disruption of AKAP anchoring decreases PKA activity in the axon.
Figure 4: Disruption of AKAP anchoring increases axon outgrowth in undifferentiated HNs.
Figure 5: Acute disruption of AKAP anchoring increases cAMP and reveals an axon-directed cAMP gradient in developing HNs.

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Acknowledgements

We thank R. Hand and A. Kolodkin from Johns Hopkins School of Medicine Department of Neuroscience for consultation on various aspects of this manuscript, reagents, and microscopy training. We thank the Malinow lab and I. Hunton at the University California San Diego for use of their lab space and coordinating dissections. We thank Zhang lab members, especially E. Greenwald, for discussions. This work was supported by the UCSD Neuroscience Microscopy Shared Facility Grant P30 NS047101, NSF GRF 1232825 (to K.G.), DGIST Convergence Science Center grant 11-BD-04 (to G.V.R.), R01NS085176 and the Craig H. Neilsen Foundation to (F.-Q.Z.), and NIH R01 DK073368 (to J.Z.).

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Authors and Affiliations

  1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Kirill Gorshkov & Jin Zhang

  2. Department of Pharmacology, University of California, San Diego, La Jolla, California, USA

    Kirill Gorshkov, Sohum Mehta & Jin Zhang

  3. Department of Neuroscience, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Santosh Ramamurthy, Gabriele V Ronnett & Feng-Quan Zhou

  4. Center for Metabolism and Obesity Research, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Santosh Ramamurthy & Gabriele V Ronnett

  5. Departments of Neurology and Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Gabriele V Ronnett

  6. Department of Orthopedic Surgery, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Feng-Quan Zhou

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  1. Kirill Gorshkov
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  2. Sohum Mehta
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Contributions

K.G. and S.R. performed the experiments; K.G., S.M., and J.Z. designed the experiments and interpreted the data. K.G., S.M., and J.Z. wrote the manuscript; K.G., S.R., S.M., and J.Z. edited the manuscript. Experiments were performed in the laboratories of G.V.R. and J.Z. F.-Q.Z. and J.Z. came up with the initial concept of the paper.

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Correspondence to Jin Zhang.

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Gorshkov, K., Mehta, S., Ramamurthy, S. et al. AKAP-mediated feedback control of cAMP gradients in developing hippocampal neurons. Nat Chem Biol 13, 425–431 (2017). https://doi.org/10.1038/nchembio.2298

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