Abstract
Homeostatic plasticity is crucial for maintaining neuronal output by counteracting unrestrained changes in synaptic strength. Chronic elevation of synaptic activity by bicuculline reduces the amplitude of miniature excitatory postsynaptic currents (mEPSCs), but the underlying mechanisms of this effect remain unclear. We found that activation of EphA4 resulted in a decrease in synaptic and surface GluR1 and attenuated mEPSC amplitude through a degradation pathway that requires the ubiquitin proteasome system (UPS). Elevated synaptic activity resulted in increased tyrosine phosphorylation of EphA4, which associated with the ubiquitin ligase anaphase-promoting complex (APC) and its activator Cdh1 in neurons in a ligand-dependent manner. APCCdh1 interacted with and targeted GluR1 for proteasomal degradation in vitro, whereas depletion of Cdh1 in neurons abolished the EphA4-dependent downregulation of GluR1. Knockdown of EphA4 or Cdh1 prevented the reduction in mEPSC amplitude in neurons that was a result of chronic elevated activity. Our results define a mechanism by which EphA4 regulates homeostatic plasticity through an APCCdh1-dependent degradation pathway.
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Acknowledgements
We thank P. Bartlett for EphA4−/− mice, R. Huganir for GluR1 antibodies and the GluR1 4KR mutant expression construct, R. Malinow for the GluR1-GFP construct, J. Chamberlain for the mouse muscle cDNA library, L. Shi, W. Chau, K. Kong, X.-N. Li, W. Fang, E. Cheung, B. Butt and C. Kwong for technical assistance, and members of the Ip laboratory for discussions. This study was supported in part by the Research Grants Council of Hong Kong SAR (HKUST, 6444/06M, 661007, 661109, 1/06C and 6/CRF/08), the Area of Excellence Scheme of the University Grants Committee (AoE/B-15/01) and the Hong Kong Jockey Club. N.Y. Ip and K.-O. Lai were recipients of the Croucher Foundation Senior Research Fellowship and Croucher Foundation Fellowship, respectively.
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N.Y.I. supervised the project. A.K.Y.F., K.-W.H., W.-Y.F., K.-O.L. and N.Y.I. designed the experiments. K.-W.H., W.-Y.F., Y.C. and K.-O.L. conducted the majority of experiments. A.K.Y.F., K.-W.H., W.-Y.F., Y.C., K.-O.L. and N.Y.I. did the data analyses. J.X. designed and did the data analyses on the electrophysiology experiment and C.S. performed electrophysiology experiment. A.K.Y.F., K.-O.L. and N.Y.I. wrote the manuscript.
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Fu, A., Hung, KW., Fu, WY. et al. APCCdh1 mediates EphA4-dependent downregulation of AMPA receptors in homeostatic plasticity. Nat Neurosci 14, 181–189 (2011). https://doi.org/10.1038/nn.2715
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DOI: https://doi.org/10.1038/nn.2715