Analysis of Histone Modifications in Rodent Pancreatic Islets by Native Chromatin Immunoprecipitation

Methods Mol Biol. 2020:2076:199-213. doi: 10.1007/978-1-4939-9882-1_11.

Abstract

The islets of Langerhans are clusters of cells dispersed throughout the pancreas that produce several hormones essential for controlling a variety of metabolic processes, including glucose homeostasis and lipid metabolism. Studying the transcriptional control of pancreatic islet cells has important implications for understanding the mechanisms that control their normal development, as well as the pathogenesis of metabolic diseases such as diabetes. Histones represent the main protein components of the chromatin and undergo diverse covalent modifications that are very important for gene regulation. Here we describe the isolation of pancreatic islets from rodents and subsequently outline the methods used to immunoprecipitate and analyze the native chromatin obtained from these cells.

Keywords: Antibodies; Histone modifications; Islets of Langerhans; Native chromatin immunoprecipitation (NChIP); Nucleosomes.

MeSH terms

  • Animals
  • Cell Separation
  • Chromatin
  • Chromatin Immunoprecipitation*
  • Gene Expression Regulation
  • Histones / metabolism*
  • Islets of Langerhans / cytology
  • Islets of Langerhans / metabolism*
  • Mice
  • Nucleosomes
  • Protein Processing, Post-Translational*
  • Rats
  • Real-Time Polymerase Chain Reaction

Substances

  • Chromatin
  • Histones
  • Nucleosomes