Cell Proliferation Analysis Using EdU Labeling in Whole Plant and Histological Samples of Arabidopsis

Methods Mol Biol. 2016:1370:169-82. doi: 10.1007/978-1-4939-3142-2_13.

Abstract

The ability to analyze cell division in both spatial and temporal dimensions within an organism is a key requirement in developmental biology. Specialized cell types within individual organs, such as those within shoot and root apical meristems, have often been identified by differences in their rates of proliferation prior to the characterization of distinguishing molecular markers. Replication-dependent labeling of DNA is a widely used method for assaying cell proliferation. The earliest approaches used radioactive labeling with tritiated thymidine, which were later followed by immunodetection of bromodeoxyuridine (BrdU). A major advance in DNA labeling came with the use of 5-ethynyl-2'deoxyuridine (EdU) which has proven to have multiple advantages over BrdU. Here we describe the methodology for analyzing EdU labeling and retention in whole plants and histological sections of Arabidopsis.

Keywords: 5-Ethynyl-2′deoxyuridine; Cell division; Cell proliferation; DNA labeling; DNA replication; EdU; Histology.

MeSH terms

  • Arabidopsis / cytology*
  • Arabidopsis / ultrastructure
  • Cell Proliferation*
  • DNA Replication
  • DNA, Plant / analysis*
  • Deoxyuridine / analogs & derivatives*
  • Deoxyuridine / analysis
  • Meristem / ultrastructure
  • Paraffin Embedding / methods
  • Plant Cells / ultrastructure
  • Plant Roots / ultrastructure
  • Seedlings / ultrastructure
  • Staining and Labeling / methods*

Substances

  • DNA, Plant
  • 5-ethynyl-2'-deoxyuridine
  • Deoxyuridine