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Transcriptional Signatures in Response to Wheat Germ Agglutinin and Starvation in Drosophila melanogaster Larval Midgut

ABSTRACT: One function of plant lectins is to serve as defenses against herbivorous insects. The midgut is the critical site affected by dietary lectins such as wheat germ agglutinin (WGA). We observed marked cellular structural and gene expression changes in Drosophila melanogaster third-instar larval midguts from insects WGA-fed or starved. Dietary WGA caused shortening, branching, swelling, distortion and in some cases disintegration of the midgut microvilli (Mv). Starvation was accompanied by shortening of the Mv. Microarray analyses revealed that dietary WGA evolved differential expression of 62 transcripts; seven of which were also differentially expressed in starved insects. The differentially regulated gene cluster in WGA-fed larvae were associated with (i) cytoskeletal organization, (ii) immune responses, (iii) detoxification reactions, and (iv) energy metabolism. Four putative transcription factor binding motifs (TFBMs) were associated with the differentially-expressed genes. At least one of these putative TFBMs exhibited substantial similarity to MyoD, a TFBM associated with cellular structures in mammals. These results are in keeping with the hypothesis that WGA causes a starvation-like effect as well as structural changes of midgut cells of Drosophila third-instar larvae. Keywords: stress response

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE11047 | GEO | 2008/10/03

SECONDARY ACCESSION(S): PRJNA107031

REPOSITORIES: GEO

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Transcriptional Signatures in Response to Wheat Germ Agglutinin and Starvation in Drosophila melanogaster Larval Midgut

Project description:One function of plant lectins is to serve as defenses against herbivorous insects. The midgut is the critical site affected by dietary lectins such as wheat germ agglutinin (WGA). We observed marked cellular structural and gene expression changes in Drosophila melanogaster third-instar larval midguts from insects WGA-fed or starved. Dietary WGA caused shortening, branching, swelling, distortion and in some cases disintegration of the midgut microvilli (Mv). Starvation was accompanied by shortening of the Mv. Microarray analyses revealed that dietary WGA evolved differential expression of 62 transcripts; seven of which were also differentially expressed in starved insects. The differentially regulated gene cluster in WGA-fed larvae were associated with (i) cytoskeletal organization, (ii) immune responses, (iii) detoxification reactions, and (iv) energy metabolism. Four putative transcription factor binding motifs (TFBMs) were associated with the differentially-expressed genes. At least one of these putative TFBMs exhibited substantial similarity to MyoD, a TFBM associated with cellular structures in mammals. These results are in keeping with the hypothesis that WGA causes a starvation-like effect as well as structural changes of midgut cells of Drosophila third-instar larvae. Experiment Overall Design: Three treatments (control, WGA, and Starvation), and three replicates for each treatment. So there are total nine samples. no dye-swap.

2008-10-26 | E-GEOD-11047 | biostudies-arrayexpress

Influences of Bwoman-Birk Inhibitor on genomics in Drosophila melanogaster larval midguts

Project description:Bowman-Birk Inhibitor (BBI) has both insecticidal and anti-cancerous properties. It has been hypothesized that dietary BBI slows insect growth by inhibiting the catalytic activity of digestive enzymes trypsins and chyomotrypsins, resulting in the midgut having reduced access to amino acids needed for growth. In mammals, BBI was hypothesized to influence cellular energy metabolism. Thus, we tested the hypothesis that dietary BBI also impacts energy-associated pathways in the midgut of Drosophila melanogaster. We investigated the impact of dietary BBI on the following parameters in the midguts of third-instar Drosophila larvae: (i) cellular metabolites, (ii) global transcriptome response, (iii) putative transcription factor binding sites (TFBSs) associated with the differentially expressed transcripts, and (iv) epithelial cellular structure. Dietary BBI caused: (i) a reduction of cellular DHAP, glucose, and succinate; and, (ii) increased Fructuse-6-phosphate; (ii) differential expression of genes associated with the glucose and fatty acid utilization; and, (iii) a shortening of midgut epithelial microvilli, a phenomenon previously associated with glucose starvation. Additionally, fifty seven percent of the putative TFBSs associated with the differentially expressed transcripts have previously been associated with glucose and insulin activities in mammalian studies. Collectively these results support the hypothesis that dietary BBI influences energy utilization in the Drosophila midgut. Keywords: stress response

2008-10-03 | GSE11046 | GEO

Influences of Bowman-Birk Inhibitor on genomics in Drosophila melanogaster larval midguts

2008-10-26 | E-GEOD-11046 | biostudies-arrayexpress

Dietary sterols/steroids and the generalist caterpillar Helicoverpa zea: physiology, biochemistry and midgut gene expression

Project description:Sterols are essential nutrients for insects because, in contrast to mammals, no insect (or arthropod for that matter) can synthesize sterols de novo. Cholesterol is the most common sterol in insects, but it is not found in plants in large quantities; plant-feeding insects typically generate their cholesterol by metabolizing phytosterols. However, different plants species can contain different types of phytosterols, and some phytosterols are not readily converted to cholesterol. In this study we examined, using artificial diets containing single sterols, how typical (cholesterol and stigmasterol) and atypical (cholestanol and cholestanone) sterols/steroids affect the performance of a generalist caterpillar (Helicoverpa zea), restricting this analysis to midgut tissue because this is where sterol/steroid absorption occurs, and the midgut is the putative site of dietary sterol/steroid metabolism. In general, H. zea performed best on the cholesterol and stigmasterol treatments; performance was reduced on cholestanol, and was very poor on cholestanone. We compared the transcript profiles of larval guts in response to differentially suitable sterols, using the optimal sterol, cholesterol, as a control, using a two-color reference design microarray experiment. Midgut gene expression patterns differed between the treatments; relative to cholesterol, differences were lowest on the stigmasterol treatment, intermediate on the cholestanol treatment, and greatest on the cholestanone treatment. Transcriptional profiling comparing Helicoverpa zea gut tissue from third instar larvae exposed to four different dietary sterols, namely Cholesterol (CON), Cholestanol (ChStanol), Cholestan-3-one (Ch3one) and Stigmasterol (Stigma). Two-color reference design. Biological replicates: 4 (5 individuals per replicate). 12 samples total.

2012-06-13 | E-GEOD-38699 | biostudies-arrayexpress

Expression profiling of HNF4-dependent transcripts in Drosophila larvae

2009-01-24 | GSE14531 | GEO

Transcription profiling of Drosophila dHNF4 null mutant larvae fed or starved for 24h

Project description:We show that dHNF4 null mutant larvae are sensitive to starvation. Starved mutant larvae consume glycogen normally, but retain lipids in their midgut and fat body, and have increased levels of long chain fatty acids, suggesting that they are unable to efficiently mobilize stored fat for energy. HNF4 dependent transcripts were identified under fed and starved conditions. Microarray studies indicate reduced expression of genes that control lipid catabolism and ï¢-oxidation. Late second instar control and HNF4 mutant larvae were collected and fed or starved for 24 hours, and then RNA was isolated for analysis. Taken together, our results support a feed-forward model for dHNF4, in which fatty acids released from triglycerides activate the receptor, inducing enzymes that drive fatty acid oxidation for energy production. Experiment Overall Design: Staged late second instar control larvae that were transheterozygous for precise excisions of the EP2449 and KG08976 P-elements, and dHNF4D33/dHNF4D17 mutant larvae, were either fed or starved for 24 hrs in a 55mm petri dish containing Whatman paper soaked with PBS. RNA was extracted from the apparently healthy animals that molted to the third instar using Trizol (Gibco) and purified on RNAeasy columns (Qiagen). All samples were prepared in triplicate to facilitate subsequent statistical analysis. Probe labeling, hybridization to Affymetrix GeneChipÂ® Drosophila Genome 2.0 Arrays.

2009-01-31 | E-GEOD-14531 | biostudies-arrayexpress

Expression profile of third instar larval fat body and midgut tissues

2010-11-02 | GSE25025 | GEO

Proteomic analysis of trachea and midgut from larvae of Drosophila melanogaster

Project description:Total protein was isolated from the trachea and midgut of D. melanogaster 3rd instar larvae and analysed by shotgun proteomics in order to identify the putative carbohydrate-active enzymes.

2018-01-11 | MSV000081913 | MassIVE

Whole genome expression analysis in the third-instar larval midgut of Drosophila melanogaster

Project description:Survival of insects on a substrate containing toxic substances such as plant secondary metabolites or insecticides is dependent on the metabolism or excretion of those xenobiotics. The primary sites of xenobiotic metabolism are the midgut, Malpighian tubules and fat body. In general, these organs are treated as single tissues by online databases, but several studies have shown that gene expression within subsections of the midgut is compartmentalized. In this article, RNA sequencing analysis was used to investigate whole-genome expression in subsections of the third-instar larval midgut. The results support functional diversification in subsections of the midgut. Analysis of the expression of gene families that are implicated in the metabolism of xenobiotics suggests that metabolism may not be uniform along the midgut. These data provide a starting point for investigating gene expression and xenobiotic metabolism in the larval midgut.

2014-09-20 | GSE32329 | GEO

Expression profile of third instar larval fat body and midgut tissues

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.

2010-11-02 | E-GEOD-25025 | biostudies-arrayexpress

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