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Mutagenesis Advance Access published online on July 14, 2007
Mutagenesis, doi:10.1093/mutage/gem026
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© The Author 2007. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org.

A comparison of G2 phase radiation-induced chromatid break kinetics using calyculin-PCC with those obtained using colcemid block

Peter E. Bryant* and Hossein Mozdarani1

Bute Medical School, University of St Andrews, St Andrews KY16 9TS, UK 1Department of Medical Genetics, School of Medical Sciences, Tarbiat Modares University, PO Box:14115-111, Tehran, Islamic Republic of Iran

To study the possible influence of cell-cycle delay on cells reaching mitosis during conventional radiation-induced chromatid break experiments using colcemid as a blocking agent, we have compared the chromatid break kinetics following a single dose of gamma rays (0.75 Gy) in metaphase CHO cells using calyculin-induced premature chromosome condensation (PCC), with those using colcemid block. Calyculin-induced PCC causes very rapid condensation of G2 cell chromosomes without the need for a cell to progress to mitosis, hence eliminating any effect of cell-cycle checkpoint on chromatid break frequency. We found that the kinetics of the exponential first-order decrease in chromatid breaks with time after irradiation was similar (not significantly different) between the two methods of chromosome condensation. However, use of the calyculin-PCC technique resulted in a slightly increased rate of disappearance of chromatid breaks and thus higher frequencies of breaks at 1.5 and 2.5 h following irradiation. We also report on the effect of the nucleoside analogue ara A on chromatid break kinetics using the two chromosome condensation techniques. Ara A treatment of cells abrogated the decrease in chromatid breaks with time, both using the calyculin-PCC and colcemid methods. We conclude that cell-cycle delay may be a factor determining the absolute frequency of chromatid breaks at various times following irradiation of cells in G2 phase but that the first-order disappearance of chromatid breaks with time and its abrogation by ara A are not significantly influenced by the G2 checkpoint.

* To whom correspondence should be addressed. Tel: +01334 463 510; Fax: +01334 463 482; Email: peb{at}st-and.ac.uk

Received on March 16, 2007; revised on May 22, 2007; accepted on May 30, 2007.


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O. Shovman, A. C. Riches, D. Adamson, and P. E. Bryant
An improved assay for radiation-induced chromatid breaks using a colcemid block and calyculin-induced PCC combination
Mutagenesis, July 1, 2008; 23(4): 267 - 270.
[Abstract] [Full Text] [PDF]


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