Adaptation to alkylation damage in DNA measured by the comet assay

Angelis, Karel J.; McGuffie, Monika; Menke, Merten; Schubert, Ingo

doi:10.1002/1098-2280(2000)36:2<146::aid-em9>3.0.co;2-5

Cited by 72 publications

(24 citation statements)

References 25 publications

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“…DSBs cause comet formation even under completely neutral conditions (N/N protocol). Treatment of DNA after lysis with specific endonucleases can be used for the selective detection of abasic sites or pyrimidine dimers [7]. Since its first application to Vicia faba [8], the comet assay has also been applied to onion [9], tobacco [10], carrot [11], Impatiens balsamina [12] and barley [13].…”

Section: Introductionmentioning

confidence: 62%

“…In this study, the A/N comet assay protocol was used. Furthermore, it was shown that adaptation to genotoxic treatments, similar to the phenomenon of clastogenic adaptation [21], can be monitored in V. faba root tips by the comet assay [7]. During adaptation, AP sites were induced, indicating that at least part of the alkyl lesions caused by MMS and MNU were removed by base excision repair.…”

Section: Monofunctional Alkylating Agents (Mnu Mms)mentioning

confidence: 99%

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DNA damage and repair in Arabidopsis thaliana as measured by the comet assay after treatment with different classes of genotoxins

Menke

Chen

Angelis

et al. 2001

Mutation Research/Genetic Toxicology and Environmental Mutagene

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143

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The three protocols of the comet assay A/N, A/A and N/N were for the first time applied to the plant species Arabidopsis thaliana. The purpose of the experiments was to establish conditions for genotoxic exposure causing DNA damage in Arabidopsis nuclei. This is required for comprehensive gene expression profiling with the intention to screen for genes involved in response of Arabidopsis cells to genotoxic stress.Five chemicals belonging to different classes of mutagens (the monofunctional alkylating agents N-methyl-N-nitrosourea and methyl methanesulfonate, the polyfunctional alkylating agent mitomycin C, the radiomimetic bleomycin and the herbicide maleic hydrazide) were tested. Except for maleic hydrazide, dose-dependent increases in DNA damage were found using the A/N comet assay protocol. While a rapid repair of bleomycin-mediated SSBs and DSBs was found, no significant reduction of DNA migration was observed up to 48 h after treatment with the monofunctional alkylating agents.

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Section: Introductionmentioning

confidence: 62%

Section: Monofunctional Alkylating Agents (Mnu Mms)mentioning

confidence: 99%

DNA damage and repair in Arabidopsis thaliana as measured by the comet assay after treatment with different classes of genotoxins

Menke

Chen

Angelis

et al. 2001

Mutation Research/Genetic Toxicology and Environmental Mutagene

Self Cite

143

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“…Ionizing radiation induces a number of DNA lesions, including oxidative base damage, single-strand breaks, and DSBs (Thacker, 1999). The methylated bases produced by MMS are repaired by enzymes of the base excision repair pathway, producing abasic sites (Angelis et al, 2000). These abasic sites inhibit DNA replication and are presumed to cause DNA Total RNA was extracted from 5-day-old seedlings untreated (C) or 30 min, 1 h, 2 h, 4 h, and 6 h after treatment with 100 Gy of irradiation.…”

Section: Atm Plants Are Hypersensitive To Ionizing Radiation and Mmsmentioning

confidence: 99%

AtATMIs Essential for Meiosis and the Somatic Response to DNA Damage in Plants[W]

et al. 2003

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In contrast to yeast or mammalian cells, little is known about the signaling responses to DNA damage in plants. We previously characterized AtATM , an Arabidopsis homolog of the human ATM gene, which is mutated in ataxia telangiectasia, a chromosome instability disorder. The Atm protein is a protein kinase whose activity is induced by DNA damage, particularly DNA double-strand breaks. The phosphorylation targets of Atm include proteins involved in DNA repair, cell cycle control, and apoptosis. Here, we describe the isolation and functional characterization of two Arabidopsis mutants carrying a T-DNA insertion in AtATM . Arabidopsis atm mutants are hypersensitive to ␥ -radiation and methylmethane sulfonate but not to UV-B light. In correlation with the radiation sensitivity, atm mutants failed to induce the transcription of genes involved in the repair and/or detection of DNA breaks upon irradiation. In addition, atm mutants are partially sterile, and we show that this effect is attributable to abundant chromosomal fragmentation during meiosis. Interestingly, the transcription of DNA recombination genes during meiosis was not dependent on AtATM , and meiotic recombination occurred at the same rate as in wild-type plants, raising questions about the function of AtAtm during meiosis in plants. Our results demonstrate that AtATM plays a central role in the response to both stress-induced and developmentally programmed DNA damage.

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“…This study was approved by the Research Committee of Hamadan University of Medical Sciences (Iran). A routine semen analysis was performed according to the World Health Organization (WHO) guidelines [17]. The sperm morphology was assessed by using the WHO criteria at a cut-off point of 30% normal sperm.…”

Section: Semen Analysismentioning

confidence: 99%

Comparison of the DNA Fragmentation and the Sperm Parameters after Processing by the Density Gradient and the Swim up Methods

Amiri¹

2012

JCDR

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Introduction:The swim up and the density-gradient centrifugation are the two main techniques which are used to separate the viable motile sperm fraction in the assisted reproductive technology. However, there are several published studies about these methods, but there is no sufficient evidence for recommending the superiority of one of them. This study was designed to study the efficiency of the swim-up and the density gradient techniques to recover the spermatozoa with a high degree of motility, a normal morphology and a low level of DNA fragmentation. Material and Methods:A total of 35 semen samples were included in the study. The semen samples were collected, one part of the semen was spread on a slide and the remainder was prepared by using the swim-up or the density gradient techniques. The recovered spermatozoa were evaluated for concentration, motility, and normal morphology. A comet assay was carried out to assay the DNA fragmentation in all the samples.Results: There were significant differences in the sperm parameters between the density gradient and the swim up techniques. Also, the swim-up technique showed a significantly higher level of DNA fragmentation as compare to the density gradient technique. Conclusion:The results of this study demonstrated several benefits of the gradients method in the separation of normal and motile spermatozoa with healthy DNA, in comparison to the swim up method.

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Adaptation to alkylation damage in DNA measured by the comet assay

Cited by 72 publications

References 25 publications

DNA damage and repair in Arabidopsis thaliana as measured by the comet assay after treatment with different classes of genotoxins

DNA damage and repair in Arabidopsis thaliana as measured by the comet assay after treatment with different classes of genotoxins

AtATMIs Essential for Meiosis and the Somatic Response to DNA Damage in Plants[W]

Comparison of the DNA Fragmentation and the Sperm Parameters after Processing by the Density Gradient and the Swim up Methods

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