A modified alkaline Comet assay for in vivo detection of oxidative DNA damage in Drosophila melanogaster

Shukla, Arvind Kumar; Pragya, Prakash; Chowdhuri, D. Kar

doi:10.1016/j.mrgentox.2011.09.017

Cited by 48 publications

(19 citation statements)

References 68 publications

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“…Oxidative damage to DNA can result in a number of different base oxidation states, which might have an impact on disease progression (cancer, cell death, carcinogenesis, and inflammation) (Shukla and others ). In the previous studies, DNA damage was assessed using comet assay, which detected the break up of DNA strand in each cell caused by free radicals (Johnson and Loo ; Guetens and others ).…”

Section: Resultsmentioning

confidence: 99%

Rosemary Extract‐Mediated Lifespan Extension and Attenuated Oxidative Damage in Drosophila melanogaster Fed on High‐Fat Diet

Wang

Sun

Rehman

et al. 2017

Journal of Food Science

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Rosemary extract has a potent antioxidant activity and is widely used in the food industry. In this study, the lifespan prolonging and antioxidant activity of rosemary extract was evaluated by high-fat-induced oxidative damage in Drosophila melanogaster. The results revealed that the lifespan and climbing ability of fruit flies was enhanced significantly by feeding rosemary extract. Furthermore, feeding with rosemary extract significantly increased the enzyme activity of superoxide dismutase (SOD) and catalase (CAT), and significantly decreased the level of malonaldehyde. The gene expression of SOD, CAT, and nuclear factor erythroid-2 related factor 2 was enhanced and that for methuselah was significantly reduced. The comet assay showed that high-fat diet-induced DNA lesion was significantly reduced in larvae treated with the rosemary extract. Our results suggest that feeding with rosemary extract is effective to the extended lifespan in fruit flies by strengthening of the resistance to high-fat-induced oxidative stress and by stimulating, at least in part, the endogenous antioxidant response.

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Section: Resultsmentioning

confidence: 99%

Rosemary Extract‐Mediated Lifespan Extension and Attenuated Oxidative Damage in Drosophila melanogaster Fed on High‐Fat Diet

Wang

Sun

Rehman

et al. 2017

Journal of Food Science

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“…In fact, non-genotoxic effects of CdCl 2 in Drosophila using the wing-spot test were observed early by our group (Rizki et al, 2004). Indirect genotoxicity of QDs in Drosophila has already been determined by means of the tunnel assay (Galeone et al, 2012) and positive effects of CdCl 2 have been reported in midgut cells (Shukla et al, 2011) and in hemocytes of spiders fed with Drosophila treated with CdCl 2 (Stalmach et al, 2014). No genotoxic effects of Cd QDs were observed in bacteria but positive induction was detected in CHO cells by means of the comet and micronucleus assays (Aye et al, 2013).…”

Section: Hsp70mentioning

confidence: 95%

Assessing potential harmful effects of CdSe quantum dots by using Drosophila melanogaster as in vivo model

Alaraby

Demir

Hernández

et al. 2015

Science of The Total Environment

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“…Cells were enzymatically individualized, incubating 15 min with collagenase (0.5 mg/mL) in PBS. Treatment times varied from 12 to 74 h ( Table 1 ; Mukhopadhyay et al, 2004; Siddique et al, 2005a,b, 2008, 2013; Mishra et al, 2011, 2013, 2014; Sharma et al, 2011, 2012; Shukla et al, 2011). …”

Section: Insightsmentioning

confidence: 99%

“…Midgut cells, with or without brain cells, have been used to study oxidative DNA damage, using incubations with FPG and Endo III enzymes (Shukla et al, 2011; Sharma et al, 2012), and to demonstrate the genotoxicity of chromium salts (Mishra et al, 2011, 2013; Sharma et al, 2011), pesticides like cypermethrin (Mukhopadhyay et al, 2004), endosulfan (Sharma et al, 2012), and dichlorvos (Mishra et al, 2014), contaminants as industrial waste leachates (Siddique et al, 2005b, 2008), and nanomaterials like graphene copper nanocomposite (Siddique et al, 2013). In addition, some of these genotoxic agents, like chromium salts, dichlorvos, and industrial waste leachates, were analyzed in different repair conditions, with the in vivo comet repair assay (Siddique et al, 2008; Mishra et al, 2011, 2013, 2014), checking the influence of pre- and post-replication DNA repair pathways on their genotoxicity.…”

Section: Usesmentioning

confidence: 99%

“…Additionally, the modified comet assay performed incubating with repair lesion-specific enzymes, as used by Shukla et al (2011) and Sharma et al (2012) for oxidative damage, can be extended to other types of damages and repair systems (Collins et al, 2008). This standardization would also clearly help the use of this assay in other types of studies, different from genotoxicity and DNA repair testing.…”

Section: Future Perspectivesmentioning

confidence: 99%

See 1 more Smart Citation

Drosophila comet assay: insights, uses, and future perspectives

Gaivão

Sierra

2014

Front. Genet.

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The comet assay, a very useful tool in genotoxicity and DNA repair testing, is being applied to Drosophila melanogaster since around 15 years ago, by several research groups. This organism is a valuable model for all kind of processes related to human health, including DNA damage response. The assay has been performed mainly in vivo using different larvae cell types (from brain, midgut, hemolymph, and imaginal disk), but also in vitro with the S2 cell line. Since its first application, it has been used to analyze the genotoxicity and action mechanisms of different chemicals, demonstrating good sensitivity and proving its usefulness. Moreover, it is the only assay that can be used to analyze DNA repair in somatic cells in vivo, comparing the effects of chemicals in different repair strains, and to quantitate repair activities in vitro. Additionally, the comet assay in Drosophila, in vivo and in vitro, has been applied to study the influence of protein overexpression on genome integrity and degradation. Although the assay is well established, it could benefit from some research to determine optimal experimental design to standardize it, and then to allow comparisons among laboratories independently of the chosen cell type.

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A modified alkaline Comet assay for in vivo detection of oxidative DNA damage in Drosophila melanogaster

Cited by 48 publications

References 68 publications

Rosemary Extract‐Mediated Lifespan Extension and Attenuated Oxidative Damage in Drosophila melanogaster Fed on High‐Fat Diet

Rosemary Extract‐Mediated Lifespan Extension and Attenuated Oxidative Damage in Drosophila melanogaster Fed on High‐Fat Diet

Assessing potential harmful effects of CdSe quantum dots by using Drosophila melanogaster as in vivo model

Drosophila comet assay: insights, uses, and future perspectives

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