title: Expression and Dendritic Trafficking of BDNF-6 Splice Variant are Impaired in Knock-In Mice Carrying Human BDNF Val66Met Polymorphism.
abstract: BACKGROUND
  The human Val66Met polymorphism in brain-derived neurotrophic factor (BDNF), a key factor in neuroplasticity, synaptic function, and cognition, has been implicated in the pathophysiology of neuropsychiatric and neurodegenerative disorders. BDNF is encoded by multiple transcripts with distinct regulation and localization, but the impact of the Val66Met polymorphism on BDNF regulation remains unclear.
  
  
  METHODS
  In BDNF Val66Met knock-in mice, which recapitulate the phenotypic hallmarks of individuals carrying the BDNF(Met) allele, we measured expression levels, epigenetic changes at promoters, and dendritic trafficking of distinct BDNF transcripts using quantitative PCR, chromatin immunoprecipitation (ChIP), and in situ hybridization.
  
  
  RESULTS
  BDNF-4 and BDNF-6 transcripts were reduced in BDNF(Met/Met) mice, compared with BDNF(Val/Val) mice. ChIP for acetyl-histone H3, a marker of active gene transcription, and trimethyl-histone-H3-Lys27 (H3K27me3), a marker of gene repression, showed higher H3K27me3 binding to exon 5, 6, and 8 promoters in BDNF(Met/Met). The H3K27 methyltransferase enhancer of zeste homolog 2 (EZH2) is involved in epigenetic regulation of BDNF expression, because in neuroblastoma cells BDNF expression was increased both by short interference RNA for EZH2 and incubation with 3-deazaneplanocin A, an inhibitor of EZH2. In situ hybridization for BDNF-2, BDNF-4, and BDNF-6 after pilocarpine treatment showed that BDNF-6 transcript was virtually absent from distal dendrites of the CA1 and CA3 regions in BDNF(Met/Met) mice, while no changes were found for BDNF-2 and BDNF-4.
  
  
  CONCLUSIONS
  Impaired BDNF expression and dendritic targeting in BDNF(Met/Met) mice may contribute to reduced regulated secretion of BDNF at synapses, and may be a specific correlate of pathology in individuals carrying the Met allele.
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee). alessandra.mallei@unimi.it.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Mallei
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee).
authors.forename: Gabriele
authors.initials: G
authors.lastname: Baj
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee).
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ieraci
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee).
authors.forename: Stefano
authors.initials: S
authors.lastname: Corna
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee).
authors.forename: Laura
authors.initials: L
authors.lastname: Musazzi
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee).
authors.forename: Francis S
authors.initials: FS
authors.lastname: Lee
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee).
authors.forename: Enrico
authors.initials: E
authors.lastname: Tongiorgi
authors.affiliation: Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative Diseases, Università degli Studi di Milano, Milano, Italy (Drs Mallei, Ieraci, Corna, Musazzi, and Popoli); Department of Life Sciences, BRAIN Centre for Neuroscience, University of Trieste, Trieste, Italy (Drs Baj and Tongiorgi); Department of Psychiatry, Weill Cornell Medical College Cornell University, New York, NY (Dr Lee).
authors.forename: Maurizio
authors.initials: M
authors.lastname: Popoli
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D003712:Dendrites / Q000187:drug effects / Q000378:metabolism*; D000071221:Enhancer of Zeste Homolog 2 Protein; D044127:Epigenesis, Genetic; D020869:Gene Expression Profiling; D055879:Gene Knock-In Techniques; D006801:Humans; D017403:In Situ Hybridization; D056484:Jumonji Domain-Containing Histone Demethylases / Q000378:metabolism; D008297:Male; D008822:Mice, Transgenic; D018721:Muscarinic Agonists / Q000494:pharmacology; D010862:Pilocarpine / Q000494:pharmacology; D063151:Polycomb Repressive Complex 2 / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms; D021381:Protein Transport / Q000187:drug effects
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D018721:Muscarinic Agonists; D020033:Protein Isoforms; D010862:Pilocarpine; C579455:BDNF protein, human; D056484:Jumonji Domain-Containing Histone Demethylases; C523761:Kdm6b protein, mouse; D000071221:Enhancer of Zeste Homolog 2 Protein; C498410:Ezh2 protein, mouse; D063151:Polycomb Repressive Complex 2
keywords: BDNF; Val66Met polymorphism; dendritic trafficking; epigenetics; histone