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Cdk5 regulates EphA4-mediated dendritic spine retraction through an ephexin1-dependent mechanism

Nature Neuroscience volume 10, pages 67–76 (2007)Cite this article

Abstract

The development of dendritic spines is thought to be crucial for synaptic plasticity. Dendritic spines are retracted upon Eph receptor A4 (EphA4) activation, but the mechanisms that control this process are not well understood. Here we report an important function of cyclin-dependent kinase 5 (Cdk5) in EphA4-dependent spine retraction in mice. We found that blocking Cdk5 activity inhibits ephrin-A1–triggered spine retraction and reduction of mEPSC frequency at hippocampal synapses. The activation of EphA4 resulted in the recruitment of Cdk5 to EphA4, leading to the tyrosine phosphorylation and activation of Cdk5. EphA4 and Cdk5 then enhanced the activation of ephexin1, a guanine-nucleotide exchange factor that regulates activation of the small Rho GTPase RhoA. The association between EphA4 and ephexin1 was significantly reduced in Cdk5−/− brains and Cdk5-dependent phosphorylation of ephexin1 was required for the ephrin-A1–mediated regulation of spine density. These findings suggest that ephrin-A1 promotes EphA4-dependent spine retraction through the activation of Cdk5 and ephexin1, which in turn modulates actin cytoskeletal dynamics.

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Figure 1: Inhibition of Cdk5 activity attenuates the ephrin-A1–stimulated dendritic spine retraction of pyramidal neurons in organotypic hippocampal slices.
Figure 2: EphA4 activation recruits the Cdk5-p35 complex and stimulates tyrosine phosphorylation of Cdk5.
Figure 3: Cdk5 activity regulates dendritic spine retraction in hippocampal neurons.
Figure 4: Ephrin-A1 does not reduce the spine density and mEPSC frequency in Cdk5−/− hippocampal neurons.
Figure 5: Cdk5 activity regulates ephrin-A1–stimulated EphA4 activation and RhoA activity in neurons.
Figure 6: Ephexin1 is phosphorylated by Cdk5-p35.
Figure 7: Cdk5 phosphorylation of ephexin1 regulates EphA4-dependent GEF activity.
Figure 8: Cdk5-dependent phosphorylation of ephexin1 is required for the ephrin-A1–mediated spine retraction.

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Acknowledgements

We are grateful to A. Kulkarni and T. Curran for Cdk5-null mice, L.H. Tsai for p35-null mice, S. Hisanaga for the recombinant Cdk5-p35 protein, D. Lo for the YFP construct and W. Chau, B. Butt, C. Kwong, K. Hung, K. Ho, W.W.Y. Chien and K. Gong for excellent technical assistance. The expert advice of D. Lo and P. Greer is gratefully acknowledged. We also thank Z. Cheung and M. Zhang for critical reading of the manuscript and members of the Ip laboratory for many helpful discussions. This study was supported in part by the Research Grants Council of Hong Kong (HKUST6131/02M, 6130/03M, 6119/04M, 6421/05M and HKUST 3/03C), the Area of Excellence Scheme of the University Grants Committee (AoE/B-15/01), the Hong Kong Jockey Club, US National Institute of Child Health and Human Development grant K08 HD01384, the William Randolph Hearst Fund (to M.S.), Mental Retardation Research Center grant HD18655, and US National Institutes of Health grant NS045500 (to M.E.G.). N.Y.I. is a Croucher Foundation Senior Research Fellow.

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  1. Wing-Yu Fu and Yu Chen: These authors contributed equally to this paper.

Authors and Affiliations

  1. Department of Biochemistry, Biotechnology Research Institute and Molecular Neuroscience Center, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, China

    Wing-Yu Fu, Yu Chen, Xiao-Su Zhao, Lei Shi, Kwok-On Lai, Amy K Y Fu, Michael E Greenberg & Nancy Y Ip

  2. Children's Hospital and Departments of Neurology and Neurobiology, Neurobiology Program, Harvard Medical School, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Mustafa Sahin & Jay B Bikoff

  3. Department of Physiology, Chinese University of Hong Kong Shatin, Hong Kong, China

    Wing-Ho Yung

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Supplementary information

Supplementary Fig. 1

EphA4 interacts with Cdk5 and p35. (PDF 478 kb)

Supplementary Fig. 2

Cdk5 is directly phosphorylated by EphA4. (PDF 379 kb)

Supplementary Fig. 3

Ephrin-A1 stimulates EphA4 clustering in cultured hippocampal neurons. (PDF 2246 kb)

Supplementary Fig. 4

Presence of EphA4, Cdk5.p35 and ephexin1 in postsynaptic densities. (PDF 472 kb)

Supplementary Fig. 5

Ephrin-A1 reduces spine density in cultured hippocampal neurons. (PDF 1864 kb)

Supplementary Fig. 6

Ephexin1 protein is phosphorylated by recombinant Cdk5.p35 using in vitro kinase assay. (PDF 574 kb)

Supplementary Fig. 7

Confocal images showing the expression of overexpressed ephexin1 in hippocampal slices. (PDF 1188 kb)

Supplementary Fig. 8

Model of regulation of spine morphogenesis by EphA4-Cdk5-ephexin1 signaling. (PDF 4228 kb)

Supplementary Methods (PDF 139 kb)

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Fu, WY., Chen, Y., Sahin, M. et al. Cdk5 regulates EphA4-mediated dendritic spine retraction through an ephexin1-dependent mechanism. Nat Neurosci 10, 67–76 (2007). https://doi.org/10.1038/nn1811

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