Document Detail


Using Drosophila larval imaginal discs to study low-dose radiation-induced cell cycle arrest.
MedLine Citation:
PMID:  21870287     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Under genotoxic stress, activation of cell cycle checkpoint responses leads to cell cycle arrest, which allows cells to repair DNA damage before continuing to cycle. Drosophila larval epithelial sacs, called imaginal discs, are an excellent in vivo model system for studying radiation-induced cell cycle arrest. Larval imaginal discs go into cell cycle arrest after being subjected to low-dose irradiation, are subject to easy genetic manipulation, are not crucial for survival of the organism, and can be dissected easily for further molecular or cellular analysis. In this chapter, we describe methods for assessing low-dose irradiation-induced cell cycle arrest. Mitotic cells are identified by immunofluorescence staining for the mitotic marker phosphorylated histone H3 (phospho-histone H3 or pH3). When wandering third-instar control larvae, without transgene expression, are exposed to 500 rads of X-ray or γ-ray irradiation, the number of pH3-positive cells in wing imaginal discs is reduced from hundreds before irradiation to approximately 30 after irradiation, with an equal distribution between the anterior and posterior compartments (Yan et al., 2011, FASEB J). Using the GAL4/UAS system, RNAi, cDNA, or microRNA sponge transgenes can be expressed in the posterior compartment of the wing disc using drivers such as engrailed (en)-Gal4, while the anterior compartment serves as an internal control. This approach makes it possible to do genome-wide genetic screening for molecules involved in radiation-induced cell cycle arrest.
Authors:
Shian-Jang Yan; Willis X Li
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  782     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2011  
Date Detail:
Created Date:  2011-08-26     Completed Date:  2012-01-09     Revised Date:  2013-06-27    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  93-103     Citation Subset:  IM    
Affiliation:
Department of Medicine, University of California, San Diego, La Jolla, CA, USA. Shianjang1_Yan@URMC.Rochester.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle Checkpoints / drug effects*
Drosophila / cytology*,  radiation effects*
Imaginal Discs / cytology*,  radiation effects*
Larva / cytology*,  radiation effects*
Grant Support
ID/Acronym/Agency:
R01 CA131326/CA/NCI NIH HHS; S10 RR026616/RR/NCRR NIH HHS; T32CA009363/CA/NCI NIH HHS
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