Title

A chromatin assay for human brain tissue

UMMS Affiliation

Psychiatry

Publication Date

12-11-2008

Document Type

Article

Subjects

Brain; *Brain Chemistry; Brain Diseases; Chromatin Immunoprecipitation; DNA Methylation; Humans

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Chronic neuropsychiatric illnesses such as schizophrenia, bipolar disease and autism are thought to result from a combination of genetic and environmental factors that might result in epigenetic alterations of gene expression and other molecular pathology. Traditionally, however, expression studies in postmortem brain were confined to quantification of mRNA or protein. The limitations encountered in postmortem brain research such as variabilities in autolysis time and tissue integrities are also likely to impact any studies of higher order chromatin structures. However, the nucleosomal organization of genomic DNA including DNA:core histone binding - appears to be largely preserved in representative samples provided by various brain banks. Therefore, it is possible to study the methylation pattern and other covalent modifications of the core histones at defined genomic loci in postmortem brain. Here, we present a simplified native chromatin immunoprecipitation (NChIP) protocol for frozen (never-fixed) human brain specimens. Starting with micrococcal nuclease digestion of brain homogenates, NChIP followed by qPCR can be completed within three days. The methodology presented here should be useful to elucidate epigenetic mechanisms of gene expression in normal and diseased human brain.

Rights and Permissions

Citation: J Vis Exp. 2008 Mar 21;(13). pii: 717. doi: 10.3791/717. Link to article on publisher's site

DOI of Published Version

10.3791/717

Related Resources

Link to Article in PubMed

Journal/Book/Conference Title

Journal of visualized experiments : JoVE

PubMed ID

19066569