Transcriptome analysis provides insights for understanding the adverse effects of endosulfan in Drosophila melanogaster

Indiscriminate use of agrochemicals worldwide, particularly, persistent organic pollutants (POPs), is of concern. Endosulfan, a POP, is used by various developing/developed nations and is known to adversely affect the development and the hormonal profiles of humans and animals. However, little is known about the molecular players/pathways underlying the adverse effects of endosulfan. We therefore analyzed the global gene expression changes and subsequent adverse effects of endosulfan using Drosophila. We used Drosophila melanogaster keeping in view of its well annotated genome and the wealth of genetic/molecular reagents available for this model organism. We exposed third instar larvae of D. melanogaster to endosulfan (2.0 μg mL⁻¹) for 24 h and using microarray, we identified differential expression of 256 genes in exposed organisms compared to controls. These genes are associated with cellular processes such as development, stress and immune response and metabolism. Microarray results were validated through quantitative PCR and biochemical assay on a subset of genes/proteins. Taking cues from microarray data, we analyzed the effect of endosulfan on development, emergence and survival of the organism. In exposed organisms, we observed deformities in hind-legs, reminiscent of those observed in higher organisms exposed to endosulfan. In addition, we observed delayed and/or reduced emergence in exposed organisms when compared to their respective controls. Together, our studies not only highlight the adverse effects of endosulfan on the organism but also provide an insight into the possible genetic perturbations underlying these effects, which might have potential implications to higher organisms.     

Mutagenic and recombinagenic activity of airborne particulates, PM10 and TSP, organic extracts in the Drosophila wing-spot test

The genotoxicity associated with air pollution in the city of Canoas, Rio Grande do Sul (Brazil), was assessed in November (spring) and January (summer). We applied the somatic mutation and recombination test (SMART) in Drosophila melanogaster in its standard version with normal bioactivation (ST) and in its variant with increased cytochrome P450-dependent biotransformation capacity (HB). The data indicated the genotoxicity of TSP and PM10 collected in November, in both ST and HB crosses. The genotoxic activity of the PM10 material in the spring sample was exclusively associated with the induction of mitotic recombination, whereas the TSP genetic toxicity was due to both recombinational as well as point and/or chromosomal mutation events. Considering PM10 collected in January, a positive response--100% (17.10 m3/ml) concentration--was observed in the HB cross, which was not detected in the ST cross.     

Toxicity and attraction effects produced byMorinda citrifolia fruits on theDrosophila melanogaster complex of species

Summary The ripe fruit of the Indian mulberry,Morinda citrifolia, is the host plant forDrosophila sechellia but is highly toxic for three closely related species (D. melanogaster, D. simulans, D. mauritiana). A simple bioassay is described with which a clear dose response to the fruit was found for these species. Significant differences in reactivity to the ripe fruit were found among species. Tested strains ofD. simulans andD. mauritiana adults were more sensitive to the toxic properties of the fruit thanD. melanogaster. A marked intraspecific variability was shown inD. melanogaster. Reciprocal interspecific hybridizations betweenD. sechellia andD. mauritiana suggested an autosomal dominant control of resistance. MoreoverD. melanogaster intraspecific crossings suggested the influence of an additional X-linked factor. Responses of flies toMorinda fruit in different states were tested in a T olfactometer. The less resistant strains ofDrosophila generally showed less preference for the ripe fruit.     

Learning and natal host influence host preference,handling time and sex allocation behaviour in a pupal parasitoid

The host choice and sex allocation decisions of a foraging female parasitoid will have an enormous influence on the life-history characteristics of her offspring. The pteromalid Pachycrepoideus vindemiae is a generalist idiobiont pupal parasitoid of many species of cyclorrhaphous Diptera. Wasps reared in Musca domestica were larger, had higher attack rates and greater male mating success than those reared in Drosophila melanogaster. In no-choice situations, na?ve female P. vindemiae took significantly less time to accept hosts conspecific with their natal host. Parasitoids that emerged from M. domestica pupae spent similar amounts of time ovipositing in both D. melanogaster and M. domestica. Those parasitoids that had emerged from D. melanogaster spent significantly longer attacking M. domestica pupae. The host choice behaviour of female P. vindemiae was influenced by an interaction between natal host and experience. Female P. vindemiae reared in M. domestica only showed a preference among hosts when allowed to gain experience attacking M. domestica, preferentially attacking that species. Similarly, female parasitoids reared on D. melanogaster only showed a preference among hosts when allowed to gain experience attacking D. melanogaster, again preferentially attacking that species. Wasp natal host also influenced sex allocation behaviour. While wasps from both hosts oviposited more females in the larger host, M. domestica, wasps that emerged from M. domestica had significantly more male-biased offspring sex ratios. These results indicate the importance of learning and natal host size in determining P. vindemiae attack rates, mating success, host preference and sex allocation behaviour, all critical components of parasitoid fitness. Electronic Publication     

Potential toxicity of flubendiamide in Drosophila melanogaster and associated structural alterations of its compound eye

Flubendiamide, a comparatively new insecticide, is used against lepidopteran insect pests and presumed safe for non-target Drosophila melanogaster (D. melanogaster). In this study, treatment concentrations (0.5, 1, 2 5, 10, and 20 μg/ml) of flubendiamide, lower than agricultural concentrations (rice 50μg/ml or cotton 100 μg/ml), were able to inhibit acetylcholinesterase activity in third instar larvae of D. melanogaster indicating a neurotoxic potential. In addition, larvae exposed to flubendiamide also manifested increased amounts of stress protein hsp70. The larvae expressing such stress response when allowed to emerge as adults displayed severe eye structure deformities found by scanning electron microscopy. These findings indicate a toxic potential for flubendiamide in D. melanogaster.     

Direct protein-protein interaction network for insecticide resistance based on subcellular localization analysis in Drosophila melanogaster

Abstract

In present study, we constructed the direct protein-protein interaction network of insecticide resistance based on subcellular localization analysis. Totally 177 of 528 resistance proteins were identified and they were located in 11 subcellular localizations. We further analyzed topological properties of the network and the biological characteristics of resistance proteins, such as k-core, neighborhood connectivity, instability index and aliphatic index. They can be used to predict the key proteins and potential mechanisms from macro-perspective. The problem of resistance has not been solved fundamentally, because the development of new insecticides can’t keep pace with the development speed of resistance, and the lack of understanding of molecular mechanism of resistance. As the further analysis to reduce data noise, we constructed the direct protein-protein interaction network of insecticide resistance based on subcellular localization analysis. The interaction between proteins located at the same subcellular location belongs to direct interactions, thus eliminating indirect interaction. Totally 177 of 528 resistance proteins were identified and they were located in 11 subcellular localizations. We further analyzed topological properties of the network and the biological characteristics of resistance proteins, such as k-core, neighborhood connectivity, instability index and aliphatic index. They can be used to predict the hub proteins and potential mechanisms from macro-perspective. This is the first study to explore the insecticide resistance molecular mechanism of Drosophila melanogaster based on subcellular localization analysis. It can provide the bioinformatics foundation for further understanding the mechanisms of insecticide resistance. It also provides a reference for the study of molecular mechanism of insecticide resistance of other insects.     

十字花科植物中的芥子碱对果蝇的抗衰老作用

芥子碱对Ｏ－２和ＯＨ的清除效率达５０％时的质量浓度（ρ）分别为５ｍｇＬ－１和３３０ｍｇＬ－１,４０ｍｇＬ－１的芥子碱对Ｏ－２的清除效率即可达８９．７％,证明芥子碱是一Ｏ－２与ＯＨ的清除剂．分别以ρ为１０ｍｇＬ－１和５０ｍｇＬ－１芥子碱的培养基饲喂果蝇,对果蝇的最高寿命、平均寿命及半数寿命均有一定的延长作用,其中１０ｍｇＬ－１的芥子碱组可明显延长雄性果蝇的最高寿命（１１．９０％）、平均寿命（１１．２４％）及半数寿命（１７．２１％）,两种浓度的芥子碱溶液对果蝇的平均寿命的延长作用的差异显著．芥子碱对果蝇的延寿作用有一定的性别差异,对雄蝇的效果好于雌蝇     

<Emphasis Type="Italic">Drosophila melanogaster</Emphasis> virgins are more likely to mate with strangers than familiar flies

Recent evidence shows that females of many species can discriminate against males and/or male phenotypes they have mated with previously. However, these studies have not tested whether actual mating is necessary to induce the avoidance behaviour. A preference for strangers may have evolved because it avoids multiple matings with similar genotypes. Alternatively, there may be selection against mating with familiar individuals directly. By choosing its first mate among unfamiliar individuals (which are less likely close relatives than are those encountered early in life), a virgin might disentangle some of the potential benefits of avoiding genetic incompatibility and inbreeding in the offspring from the costs of remating. In this study, we test whether Drosophila melanogaster flies bias their mate choice towards strangers according to previous, non-copulatory, experience. Based on 173 trials over 12 weeks, virgin females presented with two virgin males were 59% more likely to mate with a novel male than the one which she had been housed with for 8 h the day before. Hence we present the first report showing that a dipteran can distinguish between previously encountered and not previously encountered conspecifics.     

Effect of acute exposure of acephate on hemocyte abundance in a non-target victim Drosophila melanogaster

Organophosphate insecticide acephate was tested to investigate effects on hemocyte abundance in a non-target dipteran insect Drosophila melanogaster. For this purpose, third-instar larvae were fed on four graded concentrations (2, 4, 6, or 8 μg/ml) acephate for a period of 12 and 24 hr (acute treatment). Control groups were simultaneously maintained for comparison. Relative proportions of plasmatocytes in hemolymph smear were found to fall with increasing concentrations of the test chemical. Similar decreasing trend in population of lamellocytes was also noted after 12 and 24 hr chemical exposure. In contrast to plasmatocytes and lamellocytes, crystal cell number was found to rise with increasing pesticide concentration. Several factors like oxidative stress, apoptosis induction, and mitotic failure might be the cause of reduced plasmatocyte and lamellocyte count. The elevated number of crystal cells in hemolymph smears is directly indicative of high melanin synthesis that assists larvae to combat chemical stress, since melanin is well known for its potential to minimize physical, chemical, and pathogenic stress.     

Seminal fluid protein depletion and replenishment in the fruit fly, <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>: an ELISA-based method for tracking individual ejaculates

In many species, seminal fluid proteins (SFPs) affect female post-mating behavioral patterns, including sperm storage, egg laying, feeding, and remating. Yet, few studies have investigated the patterns of allocation, depletion, and replenishment of SFPs in male animals, despite the importance of these proteins to male and female reproductive success. To investigate such SFP dynamics, it is necessary to have a sensitive method for quantifying SFP levels in males and mated females. We developed such a method by adapting the enzyme-linked immunosorbent assay (ELISA) using anti-SFP antibodies. Here, we first use two Drosophila melanogaster SFPs (ovulin and sex peptide) to demonstrate that ELISAs provide accurate measures of SFP levels. We find that, consistent with previous data from Western blotting or immunofluorescence studies, levels of both ovulin and sex peptide decline in the mated female with time since mating, but they do so at different rates. We then use ELISAs to show that males become depleted of SFPs with repeated matings, but that previously mated males are able to transfer “virgin” levels of SFPs after 3 days of sexual inactivity. Finally, we demonstrate that ELISAs can detect SFPs from wild-caught D. melanogaster males and, thus, potentially can be used to track mating patterns in the wild. This method of measuring SFP dynamics can be used in a wide range of species to address questions related to male reproductive investment, female mating history, and variation in female post-mating behavioral changes. Norene A. Buehner and Anthony C. Fiumera contributed equally to this work.