Project description:We identified a role for low concentrations of 20-hydroxyecdysone (20E) in promoting growth of the Drosophila larval wing imaginal disc using genetics and in vitro explant culture. We found that 20E alone could promote proliferation in the absence of added insulin, a well known regulator of growth. To investigate the mechanism by which these hormones promote growth, we sequenced the transcriptomes of mid-third instar wing discs cultured in no hormone, 20E alone or insulin alone at two timepoints (4hr and 9hr). We also sequenced the transcriptomes of freshly dissected wing discs (not cultured). We found that within 4hr of culture without hormone, the expression of 1604 genes goes down and 1409 genes goes up, relative to uncultured discs. Changes in the expression of 1911 genes could be prevented, at least transiently, by the addition of either 20E or insulin. The transcriptional response to insulin was remarkably transient, decreasing by rougly 50% from 4hr to 9hr of culture. The response to 20E was smaller than that induced by insulin, but longer lasting. Insulin-induced genes are most significantly enriched in GO-terms describing DNA replication, ribosome biogenesis, translation, purine biosynthesis, and mitochondrial biogenesis. Insulin-repressed genes are enriched in genes involved in autophagy and cell death. 20E-regulated genes were also enriched for genes involved in DNA replication. Unlike insulin, however, 20E was found to promote the expression of genes in GO categories describing a variety of developmental processes. These data represent the first description of the direct transcriptional response of the wing disc to either insulin or low doses of 20E.
Project description:mRNA microarray of Drosophila wing imaginal discs or brain complexes from lgl27S3/lglE2S31 (lgl-null), scrib1/scrib2 or wild-type 3rd instar larvae RNA was isolated from 20 pairs of wing discs/brain complexes per sample (i.e 20 individuals). Samples were from 5 day AED wildtype and 9 day AED mutant tissue
Project description:Screening for binding partners of the splicing factor SmD3 and changes in interaction upon depletion of the protein Ecdysoneless (Ecd) in the nubbin domain of third-instar larval wing imaginal discs.
Project description:We investigated the effect on mRNA expression in Drosophila melanogaster wing imaginal discs following the knockdown of the 3'-5' exoribonuclease Dis3L2.
Project description:In order to analyze the global changes in gene expression resulting from induction of NetA-Fra signaling, we carried out a microarray experiment comparing Drosophila third instar wing imaginal discs in which Net+Fra had been overexpressed to age matched wild type wing imaginal discs. RNA extracted from both +NetA-Fra overexpression and wildtype third instar imaginal discs were hybridized to the Affymetrix GeneChip Drosophila Genome 2.0 .
Project description:We report here the transcriptomic analysis of Drosophila melanogaster wing imaginal discs from third instar female larvae mutant for corto (cortoL1/corto420) The reference line was the w1118 genetic background of the mutant lines.
Project description:In order to analyze the global changes in gene expression resulting from induction of NetA-Fra signaling, we carried out a microarray experiment comparing Drosophila third instar wing imaginal discs in which Net+Fra had been overexpressed to age matched wild type wing imaginal discs. RNA extracted from both +NetA-Fra overexpression and wildtype third instar imaginal discs were hybridized to the Affymetrix GeneChip Drosophila Genome 2.0 . Heat shock induced GFP-marked clones ectopically expressing NetA+Fra in larvae were generated. Controls for this study included age matched wildtype third instar wing imaginal discs bearing GFP clones which were prepared in the same manner. Total RNA was extracted from dissected +NetA-Fra vs. control third instar wing imaginal discs and hybridized to the Affymetrix GeneChip Drosophila Genome 2.0.
Project description:mRNA microarray of Drosophila wing imaginal discs or brain complexes from lgl27S3/lglE2S31 (lgl-null), scrib1/scrib2 or wild-type 3rd instar larvae RNA was isolated from 20 pairs of wing discs/brain complexes per sample (i.e 20 individuals). Samples were from 5 day AED wildtype and 9 day AED mutant tissue Samples were prepared in triplicate from lgl27S3/lglE2S31 (lgl-null) and scrib1/scrib2 samples and in duplicate from the wild-type 3rd instar larvae
Project description:The systemic response to injury in Drosophila melanogaster is characterized by the activation of specific signaling pathways that facilitate the regeneration of wounded tissue and help coordinate wound healing with organism growth. The mechanisms by which damaged tissues influence the development and function of peripheral non-injured tissues is not fully understood. Injury was induced in early third instar larvae via temperature-dependent cell death in wing imaginal discs. Microarray analysis using RNA isolated from injured and control was used to identify genes underlying the systemic injury response. We identified 150 genes which were differentially expressed in response to localized cell death in wing imaginal discs. Upregulated genes were associated biological processes including carnitine biosynthesis, signal transduction and regulation of oxidoreductase activity while terms associated with downregulated genes included wound healing, imaginal disc-derived wing hair outgrowth, and regulation of glutamatergic synaptic transmission. Pathway analysis revealed that wing disc damage led to changes in fatty acid, cysteine, and carnitine metabolism. One gene, 14-3-3ζ, which encodes a known regulator of Ras/MAPK signaling was identified as a potential regulator of transdetermination during tissue regeneration. Our results raise the possibility that immune function and cell proliferation during wing disc repair and regeneration in Drosophila may require the sulfur amino acid cysteine and its’ metabolites, taurine and glutathione, similar to what has been reported during tissue repair in mammals. Further, it seems likely that imaginal disc damage stimulates the mobilization of fatty acids to support the energetically demanding process of tissue regeneration. The roles of additional genes that are differentially regulated following imaginal disc injury remain to be elucidated.
