Genotype-by-diet interaction effects on transcriptome profiles in Drosophila melanogaster
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ABSTRACT: This study was performed by testing 21 wildtype genetic lines on four different diets to look at the transcriptomic, metabolomic, and gross phenotype variation. The diets used were Normal Cornmeal-molasses symbolized N, high sugar symbolized 4, high fat F, and control C a low sugar diet. Larvae were raised from hatching to 3rd instar on the respecitve diets at a density of 50 larvae per food vial. Larvae were then collected for metabolomics, transcriptomic, and gross phenotype measurements (e.g. trehalose concentration, triglyceride concentration). Samples for transcriptomic and metabolomic analyses were snap frozen in liquid nitrogen. 236 total samples were analyzed. In most cases the are unique technical replicates except in the cases where the line, diet, and biological replicate characterists are identical.
Project description:This study was performed by testing 21 wildtype genetic lines on four different diets to look at the transcriptomic, metabolomic, and gross phenotype variation. The diets used were Normal Cornmeal-molasses symbolized N, high sugar symbolized 4, high fat F, and control C a low sugar diet. Larvae were raised from hatching to 3rd instar on the respecitve diets at a density of 50 larvae per food vial. Larvae were then collected for metabolomics, transcriptomic, and gross phenotype measurements (e.g. trehalose concentration, triglyceride concentration). Samples for transcriptomic and metabolomic analyses were snap frozen in liquid nitrogen.
Project description:Chronic high sugar feeding induces obesity, hyperglycemia, and insulin resistance in flies and mammals. To gain insight into the mechanisms underlying this response, we profiled gene expression in chronically high sugar fed, wandering (post-prandial) third instar wild type larvae (L3). These data were compared to control-fed larvae as well as those (mid-L3) actively feeding for twelve hours on both diets. We used microarrays to detail the response of Drosophila larvae to high sugar-induced insulin resistance.
Project description:We utilized three ecologically diverse Drosophila species to explore the influence of ecological adaptation on transcriptomic responses to isocaloric diets differing in their relative proportions of protein to sugar. Drosophila melanogaster, a cosmopolitan species that breeds in decaying fruit, exemplifies individuals long exposed to a Western diet higher in sugar, while the natural diet of the cactophilic D. mojavensis, is much lower in carbohydrates. Drosophila arizonae, the sister species of D. mojavensis, is largely cactophilic, but also utilizes rotting fruits that are higher in sugars than cacti. We exposed third instar larvae for 24 hours to diets either (1) high in protein relative to sugar, (2) diets with equal amounts of protein and sugar, and (3) diets low in protein but high in sugar. As we predicted, based upon earlier interspecific studies of development and metabolism, the most extreme differences in gene expression under different dietary conditions were found in D. mojavensis followed by D. arizonae. No differential expression among diets was observed for D. melanogaster, a species that survives well under all three conditions, with little impact on its metabolism. We suggest that these three species together provide a model to examine individual and population differences in vulnerability to lifestyle-associated health problems such as metabolic syndrome and diabetes.
Project description:Drosophila melanogaster larvae reared on isocaloric diets with different protein/sugar ratios, exhibit different developmental times and the eclosed adults show different metabolite pools of glycogen and triglycerides (Matzkin et al., 2011, PMID: 21525254). To investigate the effect of larval diet on adult neurological processes at the gene expression level we performed high throughout RNA sequencing of fly heads reared in two different protein/sugar ratio diets.
Project description:We use mRNA-seq to transcriptionally profile larval fat body and midgut tissues from Drosophila third instar larvae. These data provide insights into tissue physiology and can be used to identify tissue specific transcripts. Fat bodies from wandering third instar larvae were dissected from ~50 male larvae and gonads were removed to eliminate contaminating transctips from the gonads. Larval midguts were dissected from ~50 wandering third instar larvae. Larval tissues were removed to Graces unsupplemented medium on ice prior to RNA extraction with TRIzol reagent. mRNA-seq samples were prepared from 5ug of total RNA and subject to Illumina based sequencing.
Project description:To determine the impact of quercetin on honeybee development and physiology, we conducted an RNASeq analysis of gene expression in neonate larvae exposed for three days to control “bee candy” diet (comprising sucrose and sugar syrup) or diets to which 0.1 mM or 0.25 mM quercetin was added.
Project description:We examined patterns of gene expression in two independent colonies of both M and S molecular forms of Anopheles gambiae at each of three developmental stages of interest: late larvae, sugar-fed virgin females, and gravid females. For each colony, replicates were derived from independent RNA samples extracted from different cohorts to ensure that trends were reproducible. In addition, each replicate was derived from larvae (adults) drawn from three pans (cages) to minimize the contribution of any individual pan to variation between samples. Data were obtained from a total of five biological replicates per mosquito colony.
Project description:Chronic high sugar feeding induces obesity, hyperglycemia, and insulin resistance in flies and mammals. To gain insight into the mechanisms underlying this response, we profiled gene expression in chronically high sugar fed, wandering (post-prandial) third instar wild type larvae (L3). These data were compared to control-fed larvae as well as those (mid-L3) actively feeding for twelve hours on both diets. We used microarrays to detail the response of Drosophila larvae to high sugar-induced insulin resistance. Male Canton-S third instar larvae were fed control (0.15M) or high (1M) sucrose and selected for RNA extraction and hybridization on Affymetrix microarrays. Wandering L3 were selected as those in the top half of the vial with partial blue guts to confirm that they had stopped eating the (blue) food. Mid-L3 were selected as L2, aged overnight until early L3, then transferred to fresh control or high sucrose food for 12 more hours before selection.
Project description:Transcriptomic data for 59 single embryos and larvae samples of the sponge Amphimedon queenslandica Transcriptomic data for 59 single embryos and larvae samples of the sponge Amphimedon queenslandica
Project description:Cancer cells utilize a unique form of aerobic glycolysis, called the Warburg effect, to efficiently produce the macromolecules required for proliferation. Here we show that a metabolic program related to the Warburg effect is used during normal Drosophila development and regulated by the fly ortholog of the Estrogen-Related Receptor (ERR) family of nuclear receptors. dERR null mutants die as second instar larvae with abnormally low ATP levels, diminished triacylglyceride stores, and elevated levels of circulating sugars. Metabolomic profiling revealed that the pathways affected in these mutants correspond to those used in the Warburg effect. The expression of active dERR protein in mid-embryogenesis triggers a coordinate switch in gene expression that drives a metabolic program supporting the dramatic growth that occurs during larval development. This study suggests that mammalian ERR family members may promote cancer by directing a metabolic state that supports proliferation. Drosophila larvae were staged at a mid-second instar time point and hand sorted for developmental progression. Individual pools of isogenic animals were collected for each replicate. Three replcates were assayed for each genotype. The two genotypes assayed were a control wild type strain (w1118) and a transheteroallelic combination of err mutant alleles (err1/err2). Labled RNA was then hybridized onto Affymetrix microarrays.