Intracellular and trans-synaptic regulation of glutamatergic synaptogenesis by EphB receptors.
The majority of mature excitatory synapses in the CNS are found on dendritic spines and contain AMPA- and NMDA-type glutamate receptors apposed to presynaptic specializations. EphB receptor tyrosine kinase signaling has been implicated in both NMDA-type glutamate receptor clustering and dendritic spine formation, but it remains unclear whether EphB plays a broader role in presynaptic and postsynaptic development. Here, we find that EphB2 is involved in organizing excitatory synapses through the independent activities of particular EphB2 protein domains. We demonstrate that EphB2 controls AMPA-type glutamate receptor localization through PDZ (postsynaptic density-95/Discs large/zona occludens-1) binding domain interactions and triggers presynaptic differentiation via its ephrin binding domain. Knockdown of EphB2 in dissociated neurons results in decreased functional synaptic inputs, spines, and presynaptic specializations. Mice lacking EphB1-EphB3 have reduced numbers of synapses, and defects are rescued with postnatal reexpression of EphB2 in single neurons in brain slice. These results demonstrate that EphB2 acts to control the organization of specific classes of mature glutamatergic synapses.
Kayser
Matthew S
MS
Department of Neuroscience, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
McClelland
Andrew C
AC
Hughes
Ethan G
EG
Dalva
Matthew B
MB
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D000076122:Disks Large Homolog 4 Protein; D004622:Embryo, Mammalian; D018698:Glutamic Acid / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D042541:Intracellular Space / Q000502:physiology*; D008164:Luminescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D016296:Mutagenesis / Q000502:physiology; D009474:Neurons / Q000166:cytology; D009924:Organ Culture Techniques; D018408:Patch-Clamp Techniques / Q000379:methods; D017729:Presynaptic Terminals / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D036183:Receptor, EphB2 / Q000378:metabolism; D036081:Receptors, Eph Family / Q000172:deficiency / Q000502:physiology*; D017470:Receptors, Glutamate / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000502:physiology*; D009435:Synaptic Transmission / Q000502:physiology*; D014162:Transfection / Q000379:methods
D000076122:Disks Large Homolog 4 Protein; C501449:Dlg4 protein, rat; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D017470:Receptors, Glutamate; D018698:Glutamic Acid; D036183:Receptor, EphB2; D036081:Receptors, Eph Family