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    Isolation of neuronal chromatin from brain tissue

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    Authors
    Jiang, Yan
    Matevossian, Anouch
    Huang, Hsien-Sung
    Straubhaar, Juerg R.
    Akbarian, Schahram
    Student Authors
    Yan Jiang; Hsien-Sung Huang
    UMass Chan Affiliations
    Program in Molecular Medicine
    Department of Psychiatry, Brudnick Neuropsychiatric Research Institute
    Document Type
    Journal Article
    Publication Date
    2008-04-30
    Keywords
    Adolescent; Adult; Aged; Animals; Antigens, Nuclear; Brain; Cell Nucleus; Child; Chromatin; Chromatin Immunoprecipitation; DNA Methylation; Flow Cytometry; Histones; Humans; Mice; Middle Aged; Molecular Biology; Nerve Tissue Proteins; Neurochemistry; Neurons; Recombinant Fusion Proteins
    Life Sciences
    Medicine and Health Sciences
    Neuroscience and Neurobiology
    
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    Abstract
    BACKGROUND: DNA-protein interactions in mature brain are increasingly recognized as key regulators for behavioral plasticity and neuronal dysfunction in chronic neuropsychiatric disease. However, chromatin assays typically lack single cell resolution, and therefore little is known about chromatin regulation of differentiated neuronal nuclei that reside in brain parenchyma intermingled with various types of non-neuronal cells. RESULTS: Here, we describe a protocol to selectively tag neuronal nuclei from adult brain - either by (anti-NeuN) immunolabeling or transgene-derived histone H2B-GFP fusion protein - for subsequent fluorescence-activated sorting and chromatin immunoprecipitation (ChIP). To illustrate an example, we compared histone H3 lysine 4 and 9 methylation marks at select gene promoters in neuronal, non-neuronal and unsorted chromatin from mouse forebrain and human cerebral cortex, and provide evidence for neuron-specific histone methylation signatures. CONCLUSION: With the modifications detailed in this protocol, the method can be used to collect nuclei from specific subtypes of neurons from any brain region for subsequent ChIP with native/un-fixed or crosslinked chromatin preparations. Starting with the harvest of brain tissue, ChIP-ready neuronal nuclei can be obtained within one day.
    Source
    BMC Neurosci. 2008 Apr 28;9:42. Link to article on publisher's site
    DOI
    10.1186/1471-2202-9-42
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/32942
    PubMed ID
    18442397
    Related Resources
    Link to Article in PubMed
    ae974a485f413a2113503eed53cd6c53
    10.1186/1471-2202-9-42
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    Morningside Graduate School of Biomedical Sciences Scholarly Publications

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