title: Neuron enriched nuclear proteome isolated from human brain.
abstract: The brain consists of diverse cell types including neurons, astrocytes, oligodendrocytes, and microglia. The isolation of nuclei from these distinct cell populations provides an opportunity to identify cell-type-specific nuclear proteins, histone modifications, and regulation networks that are altered with normal brain aging or neurodegenerative disease. In this study, we used a method by which intact neuronal and non-neuronal nuclei were purified from human post-mortem brain employing a modification of fluorescence activated cell sorting (FACS) termed fluorescence activated nuclei sorting (FANS). An antibody against NeuN, a neuron specific splicing factor, was used to isolate neuronal nuclei. Utilizing mass spectrometry (MS) based label-free quantitative proteomics, we identified 1755 proteins from sorted NeuN-positive and negative nuclear extracts. Approximately 20% of these proteins were significantly enriched or depleted in neuronal versus non-neuronal populations. Immunoblots of primary cultured rat neuron, astrocyte, and oligodendrocyte extracts confirmed that distinct members of the major nucleocytoplasmic structural linkage complex (LINC), nesprin-1 and nesprin-3, were differentially enriched in neurons and astrocytes, respectively. These comparative proteomic data sets also reveal a number of transcription and splicing factors that are selectively enriched in a cell-type-specific manner in human brain.
authors.affiliation: Department of Human Genetics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: Eric B
authors.initials: EB
authors.lastname: Dammer
authors.forename: Duc M
authors.initials: DM
authors.lastname: Duong
authors.forename: Ian
authors.initials: I
authors.lastname: Diner
authors.forename: Marla
authors.initials: M
authors.lastname: Gearing
authors.forename: Yue
authors.initials: Y
authors.lastname: Feng
authors.forename: James J
authors.initials: JJ
authors.lastname: Lah
authors.forename: Allan I
authors.initials: AI
authors.lastname: Levey
authors.forename: Nicholas T
authors.initials: NT
authors.lastname: Seyfried
mesh_terms: D001253:Astrocytes / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D044127:Epigenesis, Genetic / Q000235:genetics; D006801:Humans; D008840:Microfilament Proteins / Q000302:isolation & purification; D009419:Nerve Tissue Proteins / Q000302:isolation & purification; D009474:Neurons / Q000378:metabolism*; D009687:Nuclear Proteins / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods
chemical_list: D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; C415465:SYNE1 protein, human; C529565:SYNE3 protein, human