title: Characterization of brain cell nuclei with decondensed chromatin.
abstract: Although multipotent cell types have enlarged nuclei with decondensed chromatin, this property has not been exploited to enhance the characterization of neural progenitor cell (NPC) populations in the brain. We found that mouse brain cell nuclei that expressed exceptionally high levels of the pan neuronal marker NeuN/FOX3 (NeuN-High) had decondensed chromatin relative to most NeuN-Low or NeuN-Neg (negative) nuclei. Purified NeuN-High nuclei expressed significantly higher levels of transcripts encoding markers of neurogenesis, neuroplasticity, and learning and memory (ARC, BDNF, ERG1, HOMER1, NFL/NEF1, SYT1), subunits of chromatin modifying machinery (SIRT1, HDAC1, HDAC2, HDAC11, KAT2B, KAT3A, KAT3B, KAT5, DMNT1, DNMT3A, Gadd45a, Gadd45b) and markers of NPC and cell cycle activity (BRN2, FOXG1, KLF4, c-MYC, OCT4, PCNA, SHH, SOX2) relative to neuronal NeuN-Low or to mostly non-neuronal NeuN-Neg nuclei. NeuN-High nuclei expressed higher levels of HDAC1, 2, 4, and 5 proteins. The cortex, hippocampus, hypothalamus, thalamus, and nucleus accumbens contained high percentages of large decondensed NeuN-High nuclei, while the cerebellum, and pons contained very few. NeuN-High nuclei have the properties consistent with their being derived from extremely active neurons with elevated rates of chromatin modification and/or NPC-like cells with multilineage developmental potential. The further analysis of decondensed neural cell nuclei should provide novel insights into neurobiology and neurodegenerative disease.
authors.affiliation: Department of Genetics, University of Georgia, Davison Life Sciences Building, Athens, Georgia, 30602.
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.affiliation: Department of Genetics, University of Georgia, Davison Life Sciences Building, Athens, Georgia, 30602.
authors.forename: Elizabeth C
authors.initials: EC
authors.lastname: McKinney
authors.affiliation: Department of Genetics, University of Georgia, Davison Life Sciences Building, Athens, Georgia, 30602.
authors.forename: Muthugapatti M
authors.initials: MM
authors.lastname: Kandasamy
authors.affiliation: Department of Biology, Emory University, Atlanta, Georgia, 30322.
authors.forename: Alexandria L
authors.initials: AL
authors.lastname: Albert
authors.affiliation: Department of Genetics, University of Georgia, Davison Life Sciences Building, Athens, Georgia, 30602.
authors.forename: Richard B
authors.initials: RB
authors.lastname: Meagher
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D004268:DNA-Binding Proteins; D007150:Immunohistochemistry; D000090062:Kruppel-Like Factor 4; D008807:Mice, Inbred BALB C; D009419:Nerve Tissue Proteins / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D009419:Nerve Tissue Proteins; C492380:NeuN protein, mouse; D009687:Nuclear Proteins
keywords: cell-cycle; cytometry; epigenetics; nuclei; pluripotency; sorting