Comparative transcriptome analyses of the Drosophila pupal eye
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ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE160441 | GEO | 2020/10/30
REPOSITORIES: GEO
Similar Datasets
Project description:Hox genes regionalize the animal body axis by modifying complex morphogenetic and differentiation processes during development. The transformation of wings into halteres by the Hox gene Ultrabithorax (Ubx) in Drosophila melanogaster presents an excellent model system to study the transcriptional networks that control such complex developmental programmes. We have employed an inducible misexpression system to switch on Ubx in the wing epithelium at successive larval, prepupal and pupal stages, and have used microarray expression profiling to identify the primary transcriptional responses to Ubx. We find that Ubx regulates hundreds of downstream genes, mostly in a subtle manner. These targets are largely distinct at the different stages of appendage development and diversification. We have generated an experimental fly line combining the nabGal4NP3537-driver, a tub-GAL80ts transgene, and a UAS-UbxIa transgene (the control line was carrying a UAS-eGFP transgene instead). Our core microarray analysis has involved comparison of the transcriptional profile of experimental wings carrying the UAS-UbxIa transgene with that of control wings carrying the UAS-eGFP transgene. Pairwise comparisons have been carried out at three successive developmental stages, in particular at (i) the third instar larval wandering stage, about 4hrs before puparium formation at 29˚C, (ii) the prepupal stage, 6hrs after puparium formation (APF) at 29˚C, and (iii) the early pupal stage, 16hrs APF at 29˚C. Moreover, pairwise comparisons have been carried out with samples developed exclusively at 19˚C (UbxIa or eGFP expression OFF), as well as with samples collected at 16hrs after the temperature shift from 19 to 29˚C (UbxIa or eGFP expression ON). This has allowed us to distinguish the Ubx-dependent effects from the intrinsic expression differences between the fly lines used, and from the temperature-induced responses. We have carried out 4 biological replicates for each condition making a total of 48 hybridizations to Affymetrix Drosophila Genome 2.0 arrays.
2010-12-31 | E-GEOD-22354 | biostudies-arrayexpress
Project description:The goal of this gene expression profiling experiment was to identify the entire set of transcription factors expressed during late pupal wing development (~80h APF) when pigmentation genes are expressed
2013-02-01 | GSE43673 | GEO
Project description:The Drosophila genitalia develop from an imaginal disc that is patterned during the larval period and undergoes morphogenesis during the pupal period. Although the genetic hierarchy that controls sexual identity in Drosophila is well characterized, the downstream genes that effect sexually dimorphic development of the genitalia are largely unknown. We used microarrays to profile gene expression in female and male genital imaginal discs at three time points during development: late third-instar larvae (L3), pupae approximately 6 hours after puparium formation (P6) and pupae approximately 20 hours after puparium formation (P20). We identified genes that are sex-differentially expressed in the developing genital disc, including three genes encoding transcription factors that are expressed in the genital disc of one sex but not the other. Through genetic analysis, we showed how the master regulator of sexual identity, Doublesex, limits expression of each of these three genes to one sex. We also determined which genital structures fail to develop properly in the absence of each of the three genes. Drosophila female or male genital discs were dissected at the L3, P6 or P20 time points, total RNA was extracted, and labeled cDNA was produced for hybridization to Affymetrix microarrays. To aid in identifying genital discs, and thereby to avoid loss or contamination of tissue, we used animals expressing GFP in all imaginal discs (esgGAL4.B, UAS-GFP.nls/CyO). To collect discs of specific developmental ages, short-duration sex-nonspecific morphological features were used. L3 discs were dissected from wandering third-instar larvae and sexed by gonad size. For the two pupal time points, each sex-sorted larva was observed until it reached the white prepupa stage (0 hours after puparium formation, APF), which lasts 15-20 minutes. This short duration improves synchronization of the following stages. An air bubble forms mid-dorsally on the puparium at around 4 hours APF, peaks in size at 6-6.5 hours APF and disappears at 9-11 hours APF. We therefore used this morphological feature to collect P6 samples, rather than merely timing 6 hours since the white prepupa. Each animal was observed every 20 minutes from 5 hours APF, until peak bubble size was evident, at which time its genital disc was dissected. We followed a similar protocol for P20. We noted that GFP expression in an oval patch in the eye disc becomes faintly visible at around 20 hours APF. We observed each pupa every 20 minutes from 18 hours APF, until GFP appeared in the eye, at which time its genital disc was dissected. Morphologies of P6 and P20 discs collected this way showed little within-sex variation. For each combination of sex and time point, four biological replicates were performed.
2011-02-25 | E-GEOD-23344 | biostudies-arrayexpress
Project description:The goal of this gene expression profiling experiment was to identify the entire set of transcription factors expressed during late pupal wing development (~80h APF) when pigmentation genes are expressed We used Affymetrix microarrays to profile the expression of the transcription factors in late pupal wings of Oregon R flies. The staging of the flies was made by eye, based on the extent of wing pigmentation. RNA was extracted from batches of 40 flies and three biological replicates were analyzed.
2013-02-01 | E-GEOD-43673 | biostudies-arrayexpress
Project description:Comparative transcriptome analyses of the Drosophila pupal eye
| PRJNA673208 | ENA
Project description:To probe mechanistic determinants of Yki function in mitochondrial biogenesis, we conducted a genome-wide microarray experiment, and specifically compared expression patterns of genes from control (GMR gal4) and Yorkie over-expressing (GMR gal4; UAS yki) pupal eye discs. The above mentioned genotypes were grown at 29 deg C and at about 40 hrs after pupariation, the pupal eye discs were dissected. RNA was extracted from the pupal eye discs, purified and used to generate microarray probes that were hybridized to the Drosophila genome 2 arrays (Affymetrix). The Gene Chip Operating system (Affymetrix) and dCHIP program (Harvard University) were used to generate pairwise comparisons between the transcription profiles of control and Yorkie over-expressing discs.
2011-12-20 | E-GEOD-26700 | biostudies-arrayexpress
Project description:Expression profiling of Drosophila mir-8 homozygous mutant pupae at a single developmental stage (72 hours APF). Homozygous mutant and wild type pupae were collected at 72 hours APF (After Puparium Formation). Three independent collections were performed. Total RNA was extracted and amplified. Mutant and respective wildtype samples were hybridized together to custom cDNA arrays (DGC1 and 2 EST collections).
2007-10-11 | E-TABM-338 | biostudies-arrayexpress
Project description:To probe mechanistic determinants of Yki function in mitochondrial biogenesis, we conducted a genome-wide microarray experiment, and specifically compared expression patterns of genes from control (GMR gal4) and Yorkie over-expressing (GMR gal4; UAS yki) pupal eye discs.
2011-12-20 | GSE26700 | GEO
Project description:The male terminalia of Drosophila melanogaster has undergone rapid morphological change between it and its closely related species, making it a useful model for studying evolution and development. In order to identify the network components that pattern the male terminalia, we isolated and sequenced whole RNA from two timepoints during pupal development, and next performed in situ hybridization for 100 highly expressed transcription factors during pupal development.
2019-12-10 | GSE133732 | GEO
Project description:Drosophila melanogaster undergoes a complete metamorphosis, during which time the larval male and female forms transition into sexually dimorphic, reproductive adult forms. To understand this complex morphogenetic process at a molecular-genetic level, we performed whole genome microarray analyses. In somatic tissues at 48 hour After Puparium Formation (APF), 173 sex-biased transcripts that likely function downstream of the doublesex (dsx) branch of the sex determination hierarchy were identified. The mode of regulation of the sex-specific isoforms of DSX (DSX-F and DSX-M) was examined. It was determined that for most downstream targets, DSX-F and DSX-M regulate gene expression in the same manner, but that one isoform acts as a more potent regulator. Keywords: wild type; genetic modification All microarrays were dual channel with direct comparisons of male versus female or wild type versus mutant. All samples consist of whole body pupae collected at 48 hour After Puparium Formation (APF). For each experiment, four biological replicates were analyzed in a dye-swap design.
2009-02-10 | E-GEOD-11311 | biostudies-arrayexpress