Proposal of an in vivo comet assay using haemocytes of Drosophila melanogaster

Abstract

This study presents the first application of an in vivo alkaline comet assay using haemocytes of Drosophila melanogaster larvae. These cells, which play a role similar to that of mammalian blood, can be easily obtained and represent an overall exposure of the treated larvae. To validate the assay, we evaluated the response of these cells to three well-known mutagenic agents: ethyl methanesulfonate (EMS), potassium dichromate (PD), and gamma radiation (γ-irradiation). Third-instar Drosophila larvae were exposed to different concentrations of EMS (1, 2, and 4 mM) and PD (0.5, 1, and 2.5 mM) and to different doses of γ-irradiation (2, 4, and 8 Gγ). Subsequently, haemolymph was extracted from the larvae, and haemocytes were isolated by centrifugation and used in the comet assay. Haemocytes exhibited a significant dose-related increase in DNA damage, indicating that these cells are clearly sensitive to the treatments. These results suggest that the proposed in vivo comet test, using larvae haemocytes of D. melanogaster, may be a useful in vivo assay for genotoxicity assessment.

Genotoxicity of copper oxide nanoparticles in Drosophila melanogaster

“…An in vivo comet assay with hemocytes (blood cells) of D. melanogaster larvae was performed to detect DNA damage [36,37]. This assay has been shown to be highly sensitive, allowing for the detection of low levels of several kinds of DNA damage, such as double-and single-strand DNA breaks, alkali-labile sites, and incomplete repair sites [38].…”

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

“…Hemocytes are a suitable model to demonstrate oxidative stress induction (Alaraby et al, 2014) as well as innate immune response and DNA damage in vivo (Irving et al, 2005;Cherry and Silverman, 2006;Carmona et al, 2011). Our results showed that QDs increase ROS production in Drosophila hemocytes although to a lesser extent than after exposure to CdCl 2 .…”

“…All experiments were performed at 25 ± 1°C and at~60% relative humidity. D. melanogaster hemocytes were collected according to Carmona et al (2011). Ten microliters of cell samples (~10,000 cells) were carefully resuspended in 90 μL of 0.75% LMA at 37°C, mixed and dropped in triplicate on the hydrophilic surface of Gelbond film (GBF).…”

“…It is the fate of digested food and, inside it, immune phagocyte cells (hemocytes) are distributed which have a role similar to mammals' leucocytes. To detect the internalization of CdSe QDs in hemolymph and hemocytes, hemolymph samples were collected following a method previously described (Carmona et al, 2011). Briefly chilled 96 ± 2-h-old larvae were removed from food media, washed in water, and dried.…”

In vitro and in vivo assessment of the genotoxicity and antigenotoxicity of the Filipendula hexapetala and Filipendula ulmaria methanol extracts

“…Third instar larvae of D. melanogaster (74 ± 2 h) were placed in glass vials containing standard Drosophila diet, corn medium with EMS (1 mM in PBS) or with gallic acid (0.25 mM in PBS), and these groups were used as a negative and positive controls and standard (Siddique et al, 2005a;Carmona et al, 2011).…”