Isolation and quantification brain region-specific and cell subtype-specific histone (De)acetylation in cognitive neuroepigenetics

Abstract

The acetylation of histone tails, which relaxes compact chromatin structure and enhances the accessibility of DNA to regulatory proteins, has emerged as a key mechanism for regulating gene expression. These modifications in turn play critical roles in forming long-term memories. Chromatin immunoprecipitation (ChIP) experiments have enabled the identification of specific histone modifications and the genes most closely associated with active memory formation. Problematically, however, the majority of these studies analyze diverse populations of cell homogenates obtained from the gross dissection of large brain regions. The protocol outlined here uses methods to ascribe gene-specific histone modifications (via specific antibodies and RT-qPCR) to specific cell subtypes (via specific antibodies and cell sorting) in discrete memory-related brain regions (via microdissection) to more precisely identify the role of histone acetylation and deacetylation in cognitive neuroepigenetics.