Project description:The effect of xenobiotics (phenobarbital and atrazine) on the expression of Drosophila melanogaster CYP genes encoding cytochromes P450, a gene family generally associated with detoxification, was analyzed by DNA microarray hybridization and verified by real time RT-PCR in adults of both sexes. Only a small subset of the 86 CYP genes was significantly induced by the xenobiotics. Eleven CYP genes and three GST genes were significantly induced by phenobarbital, seven CYP and one GST gene were induced by atrazine. Cyp6d5, Cyp6w1, Cyp12d1 and the ecdysone-inducible Cyp6a2 were induced by both chemicals. The constitutive expression of several of the inducible genes (Cyp6a2, Cyp6a8, Cyp6d5, Cyp12d1) was higher in males than in females, and the induced level similar in both sexes. Thus, the level of induction was consistently higher in females than in males. The female-specific and hormonally-regulated yolk protein genes were significantly induced by phenobarbital in males and repressed by atrazine in females. Our results suggest that the numerous CYP genes of Drosophila respond selectively to xenobiotics, providing the fly with an adaptive response to chemically adverse environments. The xenobiotic-inducibility of some CYP genes previously associated with insecticide resistance in laboratory-selected strains (Cyp6a2, Cyp6a8, Cyp12d1) suggests that deregulation of P450 gene expression may be a facile way to achieve resistance. Our study also suggests that xenobiotic-induced changes in P450 levels can affect insect fitness by interfering with hormonally-regulated networks. Keywords: induction by phenobarbital and atrazine in adults drosophila melanogaster of both sex.
Project description:Piperonyl butoxide (PBO) is an insecticide synergist known to inhibit the activity of cytochrome P450 enzymes. PBO is currently used in many insecticide formulations, and has also been suggested as a pre-treatment in some pesticide applications. Little is known about how insects respond to PBO exposure at the gene transcription level. We have characterised the transcriptional response of the Drosophila melanogaster genome after PBO treatment, using both a custom designed “detox” microarray containing cytochrome P450 (P450), glutathione S-transferase (GST) and esterase genes, and a full genome microarray. We identify a subset of P450 and GST genes, along with additional metabolic genes, that are highly induced by PBO. The gene set is an extremely similar gene set to that induced by phenobarbital, a compound for which pre-treatment is known to confer tolerance to a range of insecticide compounds. The implications of the induction of gene families known to metabolise insecticides and the use of PBO in pest management programs are discussed. Keywords: Induction response after treatment by PBO
Project description:Modifications of metabolic pathways are important in insecticide resistance evolution. Mutations leading to changes in expression levels or substrate specificities of cytochrome P450 (P450), glutathione-S-transferase (GST) and esterase genes have been linked to many cases of resistance with the responsible enzyme being shown to utilize the insecticide as a substrate. Many studies show that the substrates of enzymes are capable of inducing the expression of those enzymes. We investigated if this was the case for insecticides and the enzymes responsible for their metabolism. The induction responses for P450s, GSTs and esterases to six different insecticides were investigated using a custom designed microarray in Drosophila melanogaster. Even though these gene families can all contribute to insecticide resistance, their induction responses by insecticides is minimal. The insecticides spinosad, diazinon, nitenpyram, lufenuron and dicyclanil did not induce any P450, GST or esterase gene expression. DDT was the only insecticide tested capable of eliciting an induction response, but only low levels for one GST and one P450. These results are in contrast to the induction responses observed for the natural plant compound caffeine and the barbituate drug phenobarbital, both of which induced a number of P450 and GST genes to high. Our results show that although insects evolve metabolic resistance to insecticides, induction does not usually have a role in survival after insecticide application, and induction studies cannot be used to predict which genes are capable of metabolizing insecticides. This has implications for managing the evolution of metabolic insecticide resistance in natural insect populations. Keywords: induction response after treatment by various compounds
Project description:Dietary phytochemicals are plant-derived bioactive compounds that can influence host physiology and gut microbial communities. However, the structural distinctiveness of phytochemicals and their modulation of host intestinal gene expression in a sex-dependent manner remain poorly understood. In this study, we used Drosophila melanogaster to investigate gut transcriptomic responses to four phytochemical-enriched diets, capsaicin, curcumin, resveratrol, and thymoquinone. Two-day-old mated female and male flies were exposed to control or phytochemical-supplemented diets for 10 days, after which gut tissues were dissected for bulk RNA sequencing. Transcriptomic analysis revealed that biological sex was a major source of variation in the gut, with female and male flies showing distinct expression profiles across dietary treatments. Phytochemical supplementation induced compound and sex-specific transcriptional responses. Curcumin produced the strongest transcriptional response, with females showing reduced expression of genes involved in carbohydrate digestion and trehalose metabolism, whereas males showed enrichment of xenobiotic detoxification pathways dominated by cytochrome P450 genes. Capsaicin also induced sex-dependent responses, including detoxification-associated transcriptional changes in females and increased expression of immune-related genes in males. These results indicate that dietary phytochemicals elicit sex-specific gut transcriptomic responses in Drosophila melanogaster and highlight biological sex as an important variable in studies of diet-host interactions.
Project description:Proteomic Analysis (MS/MS) of Drosophila melanogaster mtx2 (Ortholog of CG8004) Heterozygous versus Homozygous Mutants at 2 Days Post-Pupa Formation