Immunochemical Detection of Modified Cytosine Species in Lampbrush Chromatin

Methods Mol Biol. 2021:2198:159-168. doi: 10.1007/978-1-0716-0876-0_13.

Abstract

The lampbrush chromosomes found in the giant nucleus or germinal vesicle (GV) of amphibian oocytes provide unique opportunities for discrete closed and open chromatin structural domains to be directly observable by simple light microscopy. Moreover, the method described here for preparing spreads of lampbrush chromatin for immunostaining enables a straightforward approach to establishing the distributions of modified nucleotides within and between structurally and functionally distinctive chromatin domains.

Keywords: Chromatin domain; Chromosome; Germinal vesicle; Xenopus; axolotl.

MeSH terms

  • Animals
  • Cell Nucleus / immunology
  • Chromatin / genetics
  • Chromatin / immunology*
  • Chromosomes / immunology
  • Cytosine / chemistry
  • Cytosine / immunology
  • Female
  • Immunohistochemistry / methods*
  • Oocytes / immunology*
  • Oocytes / metabolism
  • Reptiles / embryology
  • Reptiles / immunology
  • Xenopus laevis / genetics

Substances

  • Chromatin
  • Cytosine