DNA Strand Breaks in Mitotic Germ Cells of Caenorhabditis elegans Evaluated by Comet Assay

Mol Cells. 2016 Mar;39(3):204-10. doi: 10.14348/molcells.2016.2206. Epub 2016 Feb 23.

Abstract

DNA damage responses are important for the maintenance of genome stability and the survival of organisms. Such responses are activated in the presence of DNA damage and lead to cell cycle arrest, apoptosis, and DNA repair. In Caenorhabditis elegans, double-strand breaks induced by DNA damaging agents have been detected indirectly by antibodies against DSB recognizing proteins. In this study we used a comet assay to detect DNA strand breaks and to measure the elimination of DNA strand breaks in mitotic germline nuclei of C. elegans. We found that C. elegans brc-1 mutants were more sensitive to ionizing radiation and camptothecin than the N2 wild-type strain and repaired DNA strand breaks less efficiently than N2. This study is the first demonstration of direct measurement of DNA strand breaks in mitotic germline nuclei of C. elegans. This newly developed assay can be applied to detect DNA strand breaks in different C. elegans mutants that are sensitive to DNA damaging agents.

Keywords: Caenorhabditis elegans; DNA damage; DNA repair; DNA strand break; comet assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans / drug effects
  • Caenorhabditis elegans / embryology*
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans / radiation effects
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism
  • Camptothecin / pharmacology
  • Cell Nucleus / drug effects
  • Cell Nucleus / genetics
  • Cell Nucleus / radiation effects
  • Comet Assay / methods*
  • DNA Breaks, Double-Stranded* / drug effects
  • DNA Breaks, Double-Stranded* / radiation effects
  • DNA Repair
  • Embryo, Nonmammalian / drug effects
  • Embryo, Nonmammalian / radiation effects
  • Germ Cells / cytology*
  • Germ Cells / drug effects
  • Germ Cells / radiation effects
  • Mitosis* / drug effects
  • Mitosis* / radiation effects
  • Mutation
  • Radiation, Ionizing

Substances

  • BRC-1 protein, C elegans
  • Caenorhabditis elegans Proteins
  • Camptothecin