Methods for studying the DNA damage response in the Caenorhabdatis elegans germ line.
In response to genotoxic insults, cells activate DNA damage response pathways that either stimulate transient cell cycle arrest and DNA repair or induce apoptosis. The Caenorhabditis elegans germ line is now well established as a model system to study these processes in a genetically tractable, multicellular organism. Upon treatment with genotoxic agents, premeiotic C. elegans germ cells transiently halt cell cycle progression, whereas meiotic prophase germ cells in the late-pachytene stage undergo apoptosis. Further, accumulation of unrepaired meiotic recombination intermediates can also lead to apoptosis of affected pachytene cells. DNA damage-induced cell death requires key components of the evolutionarily conserved apoptotic machinery. Moreover, both cell cycle arrest and pachytene apoptosis responses depend on conserved DNA damage checkpoint proteins. Genetics- and genomics-based approaches that have demonstrated roles for conserved checkpoint proteins have also begun to uncover novel components of these response pathways. In this chapter, we briefly review the C. elegans DNA damage response field, discuss in detail methods currently used to assay DNA damage responses in C. elegans, and describe the development of new experimental tools that will facilitate a more comprehensive understanding of the DNA damage response.
Wellcome Trust Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Scotland, United Kingdom.
Ashley L
AL
Craig
Sandra C
SC
Moser
Aymeric P
AP
Bailly
Anton
A
Gartner
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D001681:Biological Assay; D015415:Biomarkers / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage; D004260:DNA Repair; D005720:Gamma Rays; D005854:Germ Cells / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D006918:Hydroxyurea / Q000494:pharmacology; D007814:Larva / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D008540:Meiosis / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D008938:Mitosis / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D034622:RNA Interference; D015398:Signal Transduction / Q000235:genetics
D015415:Biomarkers; D029742:Caenorhabditis elegans Proteins; D006918:Hydroxyurea