GSBS Student Publications

Student Author(s)

Yan Jiang; Hsien-Sung Huang

GSBS Program

Neuroscience

UMMS Affiliation

Department of Psychiatry, Brudnick Neuropsychiatric Research Institute; Program in Molecular Medicine

Date

4-30-2008

Document Type

Article

Medical Subject Headings

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

Disciplines

Life Sciences | Medicine and Health Sciences | Neuroscience and Neurobiology

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.

Rights and Permissions

Citation: BMC Neurosci. 2008 Apr 28;9:42. Link to article on publisher's site

DOI of Published Version

10.1186/1471-2202-9-42

Related Resources

Link to Article in PubMed

Journal Title

BMC neuroscience

PubMed ID

18442397

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