title: Activity and protein kinase C regulate synaptic accumulation of N-methyl-D-aspartate (NMDA) receptors independently of GluN1 splice variant.
abstract: NMDA receptors are calcium-permeable ionotropic receptors that detect coincident glutamate binding and membrane depolarization and are essential for many forms of synaptic plasticity in the mammalian brain. The obligatory GluN1 subunit of NMDA receptors is alternatively spliced at multiple sites, generating forms that vary in N-terminal N1 and C-terminal C1, C2, and C2' cassettes. Based on expression of GluN1 constructs in heterologous cells and in wild type neurons, the prevalent view is that the C-terminal cassettes regulate synaptic accumulation and its modulation by homeostatic activity blockade and by protein kinase C (PKC). Here, we tested the role of GluN1 splicing in regulated synaptic accumulation of NMDA receptors by lentiviral expression of individual GluN1 splice variants in hippocampal neurons cultured from GluN1 (-/-) mice. High efficiency transduction of GluN1 at levels similar to endogenous was achieved. Under control conditions, the C2' cassette mediated enhanced synaptic accumulation relative to the alternate C2 cassette, whereas the presence or absence of N1 or C1 had no effect. Surprisingly all GluN1 splice variants showed >2-fold increased synaptic accumulation with chronic blockade of NMDA receptor activity. Furthermore, in this neuronal rescue system, all GluN1 splice variants were equally rapidly dispersed upon activation of PKC. These results indicate that the major mechanisms mediating homeostatic synaptic accumulation and PKC dispersal of NMDA receptors occur independently of GluN1 splice isoform.
authors.affiliation: Center for Neuroscience and Cell Biology and Department of Life Sciences, University of Coimbra, 3004-517 Coimbra, Portugal.
authors.forename: Joana S
authors.initials: JS
authors.lastname: Ferreira
authors.forename: Amanda
authors.initials: A
authors.lastname: Rooyakkers
authors.forename: Kevin
authors.initials: K
authors.lastname: She
authors.forename: Luis
authors.initials: L
authors.lastname: Ribeiro
authors.forename: Ana Luísa
authors.initials: AL
authors.lastname: Carvalho
authors.forename: Ann Marie
authors.initials: AM
authors.lastname: Craig
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D000818:Animals; D004789:Enzyme Activation / Q000502:physiology; D051379:Mice; D018345:Mice, Knockout; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D016194:Receptors, N-Methyl-D-Aspartate / Q000096:biosynthesis* / Q000235:genetics; D013570:Synaptic Membranes / Q000235:genetics / Q000378:metabolism*
chemical_list: D016194:Receptors, N-Methyl-D-Aspartate; D011493:Protein Kinase C