Serine phosphorylation of ephrinB2 regulates trafficking of synaptic AMPA receptors

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Plasticity in the brain is essential for maintaining memory and learning and is associated with the dynamic membrane trafficking of AMPA receptors. EphrinB proteins, ligands for EphB receptor tyrosine kinases, are transmembrane molecules with signaling capabilities that are required for spine morphogenesis, synapse formation and synaptic plasticity. Here, we describe a molecular mechanism for ephrinB2 function in controlling synaptic transmission. EphrinB2 signaling is critical for the stabilization of AMPA receptors at the cellular membrane. Mouse hippocampal neurons from conditional ephrinB2 knockouts showed enhanced constitutive internalization of AMPA receptors and reduced synaptic transmission. Mechanistically, glutamate receptor interacting proteins bridge ephrinB ligands and AMPA receptors. Moreover, this function involved a regulatory aspect of ephrinB reverse signaling that involves the phosphorylation of a single serine residue in their cytoplasmic tails. In summary, our findings uncover a model of cooperative AMPA receptor and ephrinB reverse signaling at the synapse.

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Figure 1: EphrinB2 reverse signaling regulates AMPA receptor endocytosis.
Figure 2: EphrinB2 knockout hippocampal neurons show enhanced constitutive internalization of AMPA receptors and reduced synaptic transmission.
Figure 3: GRIP molecules bridge ephrinB ligands and AMPA receptors.
Figure 4: GRIP molecules are required for ephrinB ligand–mediated AMPA receptor stabilization.
Figure 5: GRIP binding to ephrinBs is regulated by EphB receptors.
Figure 6: EphrinB ligands are phosphorylated on serine residues.
Figure 7: Serine phosphorylation of ephrinB ligands regulates PDZ-dependent interactions.
Figure 8: Serine phosphorylation of ephrinB ligands regulates AMPA receptor internalization.


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We would like to thank A.Yamasaki for technical help, J. Lauterbach and G. Wilkinson for valuable reagents, I. Kadow for helpful discussions, and A.Yamasaki, E. Fellows, I. Kadow, T. Acker and S. Sigrist for critically reading the manuscript. This work was supported by grants from the Deutsche Forschungsgemeinschaft (AC180/2-1 and AC180/2-2 to A.A.-P.) and the Cluster of Excellence 'Macromolecular Complexes' at the Goethe University Frankfurt (EXC 115).

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All the antibody feeding assays, surface biotinylation assays, colocalization and surface clustering experiments were carried out by C.L.E. with the help of E.M. and J.C.G. M.Z. and E.M. performed the biochemical and cellular characterization of GRIP binding to ephrinB ligands. R.K. supervised and supported the work of M.Z. Eph-GRIP–binding experiments were carried out by E.M. M.H.T. performed the electrophysiology under the supervision of V.S. A.A.-P. carried out the kinase assays, designed and supervised the experiments, and wrote the manuscript.

Correspondence to Amparo Acker-Palmer.

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Essmann, C., Martinez, E., Geiger, J. et al. Serine phosphorylation of ephrinB2 regulates trafficking of synaptic AMPA receptors. Nat Neurosci 11, 1035–1043 (2008) doi:10.1038/nn.2171

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