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Mutagenesis Advance Access published online on May 17, 2005
Mutagenesis, doi:10.1093/mutage/gei032
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© The Author 2005. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please email: journals.permissions@oupjournals.org
Received October 10, 2004
Accepted April 17, 2005

Original article

Validation of Drosophila melanogaster as an in vivo model for genotoxicity assessment using modified alkaline Comet assay

Hifzur R. Siddique 1, D. Kar Chowdhuri 1, D. K. Saxena 1, and Alok Dhawan 2*

1 Embryotoxicology Section, Industrial Toxicology Research Center, PO Box 80, M.G. Marg, Lucknow, 226001, Uttar Pradesh, India
2 Developmental Toxicology Section, Industrial Toxicology Research Center, PO Box 80, M.G. Marg, Lucknow, 226001, Uttar Pradesh, India

* To whom correspondence should be addressed.
Alok Dhawan, E-mail: dhawanalok{at}hotmail.com


  Abstract

The single cell gel electrophoresis or Comet assay is one of the most popular techniques for genotoxicity assessment. The present study was undertaken to validate our previously modified version of the Comet assay for genotoxicity assessment in Drosophila melanogaster (Oregon R+) with four well-known mutagenic and carcinogenic alkylating agents, i.e. ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), N-ethyl-N-nitrosourea (ENU) and cyclophosphamide (CP). Third instar larvae (74 ± 2 h) of D.melanogaster were fed different concentrations of EMS, MMS, ENU and CP (0.05, 0.5 and 1.0 mM) mixed standard Drosophila food for 24 h. At 98 ± 2 h, the anterior midgut from control and treated larvae were dissected out, single-cell suspensions were prepared and Comet assay was performed. Our results show a dose-dependent increase in DNA damage with all the four alkylating agents, in comparison to control. The lower concentration (0.05 mM) of the test chemicals, except MMS, did not induce any DNA damage in the gut cells of the exposed larvae. When comparison of Comet parameters was made among the chemicals, MMS was found to be the most potent genotoxicant and ENU the least. The present study validated our previous observation and shows that D.melanogaster is a sensitive and suitable model for the in vivo assessment of genotoxicity using our modified alkaline Comet assay.


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