Arabinosylcytosine-induced accumulation of DNA nicks in myotube nuclei detected by in situ nick translation. J Cell Physiol 1990 Jul;144(1):12-7
Date
07/01/1990Pubmed ID
2365739DOI
10.1002/jcp.1041440103Scopus ID
2-s2.0-0025316415 (requires institutional sign-in at Scopus site) 13 CitationsAbstract
This laboratory has recently reported the occurrence of DNA nicking at the onset of terminal skeletal myogenesis by using the technique of in situ nick translation (Dawson and Lough: Dev. Biol., 127:362-367, 1988). Because 1-beta-D-arabinofuranosylcytosine (araC), a cytocidal agent that is routinely used to removed dividing fibroblasts from myogenic cultures, inhibits DNA repair, it was of interest to determine whether araC treatment resulted in an accumulation of the endogenously created nicks. Thus, we have assessed the accumulation of DNA nicks in myotube cells during a 20 hour araC treatment period at the onset of terminal myogenesis (44-64 hours in vitro) by using three techniques: alkaline sucrose gradient density centrifugation, kinetic in situ nick translation, and cellular in situ nick translation. Although alkaline sucrose gradient centrifugation revealed no detectable nicking after 20 hours, kinetic in situ nick translation analysis revealed subtle but significant increases in DNA nicks caused by araC within 7 hours of drug application, and a 1.5-fold increase in DNA repair sites after 20 hours of drug treatment. That these observations reflected nicking specifically in myotube nuclei was determined by immunocytochemical localization of nicked sites after repair with a biotinylated nucleotide analog (biotin-11-dUTP). The effects of araC were only incompletely reversible, whether or not the drug was removed from the cultures, within 2 days of the treatment period.(ABSTRACT TRUNCATED AT 250 WORDS)
Author List
Masuck TM, Taylor AR, Lough JMESH terms used to index this publication - Major topics in bold
AnimalsCell Differentiation
Cell Nucleus
Centrifugation, Density Gradient
Chick Embryo
Cytarabine
DNA
DNA Damage
DNA Repair
In Vitro Techniques
Muscles
