title: NR2B phosphorylation at tyrosine 1472 contributes to brain injury in a rodent model of neonatal hypoxia-ischemia.
abstract: OBJECTIVE
  The NR2B subunit of the N-methyl-d-aspartate (NMDA) receptor is phosphorylated by the Src family kinase Fyn in brain, with tyrosine (Y) 1472 as the major phosphorylation site. Although Y1472 phosphorylation is important for synaptic plasticity, it is unknown whether it is involved in NMDA receptor-mediated excitotoxicity in neonatal brain hypoxia-ischemia (HI). This study was designed to elucidate the specific role of Y1472 phosphorylation of NR2B in neonatal HI in vivo and in NMDA-mediated neuronal death in vitro.
  
  
  METHODS
  Neonatal mice with a knockin mutation of Y1472 to phenylalanine (YF-KI) and their wild-type littermates were subjected to HI using the Vannucci model. Brains were scored 5 days later for damage using cresyl violet and iron staining. Western blotting and immunoprecipitation were performed to determine NR2B tyrosine phosphorylation. Expression of NADPH oxidase subunits and superoxide production were measured in vivo. NMDA-induced calcium response, superoxide formation, and cell death were evaluated in primary cortical neurons.
  
  
  RESULTS
  After neonatal HI, YF-KI mice have reduced expression of NADPH oxidase subunit gp91phox and p47phox and superoxide production, lower activity of proteases implicated in necrotic and apoptotic cell death, and less brain damage when compared with the wild-type mice. In vitro, YF-KI mutation diminishes superoxide generation in response to NMDA without effect on calcium accumulation and inhibits NMDA and glutamate-induced cell death.
  
  
  CONCLUSIONS
  Upregulation of NR2B phosphorylation at Y1472 after neonatal HI is involved in superoxide-mediated oxidative stress and contributes to brain injury.
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Renatta
authors.initials: R
authors.lastname: Knox
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Angela M
authors.initials: AM
authors.lastname: Brennan-Minnella
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Fuxin
authors.initials: F
authors.lastname: Lu
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Diana
authors.initials: D
authors.lastname: Yang
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Takanobu
authors.initials: T
authors.lastname: Nakazawa
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Tadashi
authors.initials: T
authors.lastname: Yamamoto
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Raymond A
authors.initials: RA
authors.lastname: Swanson
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.).
authors.forename: Donna M
authors.initials: DM
authors.lastname: Ferriero
authors.affiliation: From the Department of Pediatrics (R.K., F.L., D.Y., D.M.F., X.J.), Biomedical Sciences Graduate Program (R.K., D.M.F.), Medical Scientist Training Program (R.K.), Department of Neurology (A.M.B.-M., R.A.S., D.M.F.), and San Francisco Veterans Affairs Medical Center (A.M.B.-M., R.A.S.), University of California, San Francisco; and Division of Oncology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan (T.N., T.Y.). xiangning.jiang@ucsf.edu.
authors.forename: Xiangning
authors.initials: X
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D004195:Disease Models, Animal; D005455:Fluorescent Antibody Technique; D055879:Gene Knock-In Techniques; D020925:Hypoxia-Ischemia, Brain / Q000378:metabolism* / Q000473:pathology*; D047468:Immunoprecipitation; D051379:Mice; D008810:Mice, Inbred C57BL; D018384:Oxidative Stress; D010766:Phosphorylation; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
chemical_list: C121001:NR2B NMDA receptor; D016194:Receptors, N-Methyl-D-Aspartate; D014443:Tyrosine
keywords: Fyn tyrosine kinase; NR2B NMDA receptor; hypoxia-ischemia, brain