Visualization of HDAC9 Spatiotemporal Subcellular Localization in Primary Neuron Cultures

Methods Mol Biol. 2016:1436:119-27. doi: 10.1007/978-1-4939-3667-0_9.

Abstract

Histone deacetylase (HDAC) 9 is one of class IIa HDACs which are expressed in developing cortical neurons. The translocation of HDAC9 from the nucleus to the cytoplasm is induced by neuronal activity during postnatal development, and is involved in regulation of various gene expressions. Visualization of HDAC9 subcellular localization is a powerful tool for studying activity-dependent gene expression. Here, we describe a time-lapse imaging method using fluorescent protein-tagged HDAC9 in dissociated cortical neurons. This method reveals dynamic HDAC9-mediated gene expression in response to various signals.

Keywords: Fluorescent imaging; HDAC9; Primary cortical neuron cultures; Time-lapse imaging; Transfection; cDNA cloning.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Nucleus / metabolism*
  • Cells, Cultured
  • Cloning, Molecular
  • Cytoplasm / metabolism*
  • Gene Expression Regulation
  • Histone Deacetylases / genetics
  • Histone Deacetylases / metabolism*
  • Mice
  • Microscopy, Fluorescence
  • Neurons / cytology*
  • Neurons / metabolism
  • Neurons / ultrastructure
  • Protein Transport
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Time-Lapse Imaging / methods*

Substances

  • Repressor Proteins
  • Hdac9 protein, mouse
  • Histone Deacetylases