Genome-wide expression profiling in Drosophila wing imaginal discs
Ontology highlight
ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE180120 | GEO | 2021/07/15
REPOSITORIES: GEO
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Project description:Signaling between cells in the Anterior (A) and Posterior (P) compartments directs Drosophila wing disc development and is dependent on expression of the homeodomain transcription factor Engrailed (En) in P cells. Downstream of en, posteriorly expressed Hedgehog (Hh) protein signals across the A/P border to establish a developmental organizer that directs pattern formation and growth throughout the wing primordium. Here we extend investigations of the processes downstream of en by using expression array analysis to compare A and P cells. 102 candidate genes were identified that express differentially in the A and P compartments; four were characterized: Stubble (Sb) expression is restricted to A cells due to repression by en. CG15905, CG16884, and CG10200/hase und igel (hui) are expressed in A cells downstream of Hh signaling; and RNA interference for hui, Stubble, and CG16884 revealed that each is essential to wing development. We carried out a global screen for genes with compartment-specific expression using expression array hybridization to compare transcript levels in A and P wing disc cells. GFP-labeled wing imaginal discs (ptc-Gal4/UAS-GFP and hh-Gal4/UAS-GFP) were micro-dissected under a fluorescence dissecting microscope. RNA isolation, amplification and microarray procedures were previously described (Klebes et al., 2002; Klebes et al., 2005; Klebes and Kornberg, 2008). In brief, hybridization probes were generated by two rounds of T7-catalyzed linear RNA amplification and labeled with Cy3 and Cy5 dyes. Reciprocally labeled probes (M-bM-^@M-^Xdye flipM-bM-^@M-^Y) were hybridized to custom produced glass microarrays (GPL2581) that contained approximately 14,000 100-600 bp exon sequences that were generated by PCR. Signal intensities were collected with a GenePix 4000B Scanner and processed with GenePix software (Molecular Devices) and global median normalized with NOMAD (http://ucsf-nomad.sourceforge.net/).
2013-05-04 | E-GEOD-46601 | biostudies-arrayexpress
Project description:The patterning of Drosophila retina occurs both very fast and with high precision. This process is driven by the dynamic changes in signalling activity of the conserved Hedgehog (Hh) pathway, which coordinates cell fate determination, cell cycle and tissue morphogenesis. Here we show that during Drosophila retinogenesis, the retinal determination gene dachshund (dac) is not only a target of the Hh signaling pathway, but is also a modulator of its activity. Using developmental genetics techniques, we demonstrate that dac enhances Hh signaling by promoting the accumulation of the Gli transcription factor Cubitus interruptus (Ci) parallel to or downstream of fused. In the absence of dac, all Hh-mediated events associated to the morphogenetic furrow are delayed. One of the consequences is that, posterior to the furrow, dac- cells cannot activate a Roadkill-Cullin3 negative feedback loop that attenuates Hh signaling and which is necessary for retinal cells to continue normal differentiation. Therefore, dac is part of an essential positive feedback loop in the Hh pathway, guaranteeing the speed and the accuracy of Drosophila retinogenesis. ChIP-seq against Dachshund vs input ChIP-seq. Eye-antennal imaginal discs are dissected from Grh-GFP (Bloomington stock 42269) 3rd instar larvae and fixed with formaldehyde. Chromatin is prepared and sonicated until fragments reach an average size of 500 bp. Chromatin is immunoprecipitated with an anti-GFP Ab (ab290, Abcam) and the immunocomplexes are recovered with protein A/G magnetic beads (Millipore).
2016-07-01 | E-GEOD-82151 | biostudies-arrayexpress
Project description:In the spider Parasteatoda tepidariorum, Hedgehog (Hh) and Decapentaplegic (Dpp) signaling play key roles in the formation of the two major embryonic axes (Akiyama-Oda and Oda, 2010, Development 137, 1263-1273; Akiyama-Oda and Oda, 2006, Development 133, 2347-2357). Patched (Ptc), a receptor for Hh,functions as a negative regulator of Hh signaling. Double parental RNA interference (pRNAi) against spider homologs of ptc and dpp, Pt-ptc and Pt-dpp, respectively, results in severe defects in axis formation. In this microarray experiment, we attempted to identify candidate genes whose expressions are regulated by Hh and Dpp signaling during axis formation.
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Project description:Signaling between cells in the Anterior (A) and Posterior (P) compartments directs Drosophila wing disc development and is dependent on expression of the homeodomain transcription factor Engrailed (En) in P cells. Downstream of en, posteriorly expressed Hedgehog (Hh) protein signals across the A/P border to establish a developmental organizer that directs pattern formation and growth throughout the wing primordium. Here we extend investigations of the processes downstream of en by using expression array analysis to compare A and P cells. 102 candidate genes were identified that express differentially in the A and P compartments; four were characterized: Stubble (Sb) expression is restricted to A cells due to repression by en. CG15905, CG16884, and CG10200/hase und igel (hui) are expressed in A cells downstream of Hh signaling; and RNA interference for hui, Stubble, and CG16884 revealed that each is essential to wing development. We carried out a global screen for genes with compartment-specific expression using expression array hybridization to compare transcript levels in A and P wing disc cells.
2013-05-04 | GSE46601 | GEO
Project description:Hedgehog signaling plays pivotal roles in formation of the embryonic anterior-posterior axis in the spider Parasteatoda tepidariorum. In this work, using the combination of RNA-sequencing and parental RNA interference (pRNAi), we genome-widely identified genes that transcriptionally responded to altered states of Hedgehog signaling in spider embryos, which were caused by Pt-hedgehog (Pt-hh) pRNAi and Pt-patched (Pt-ptc) pRNAi. The identified genes were classified into four classes (Classes I to IV) based on the positive/negative responses to the Pt-hh pRNAi and Pt-ptc pRNAi states. The Class II genes were negatively regulated by Hh signaling (positive response to Pt-hh pRNAi and negative response to Pt-ptc pRNAi). Among the Class II genes, we identified Pt-msx1 as a key segmentation gene in the spider embryo. Moreover, we conducted a transcriptomic analysis of Pt-msx1 pRNAi embryos and identified additional genes whose expression showed patterns associated with segmentation.
2020-04-01 | GSE136357 | GEO
Project description:The patterning of Drosophila retina occurs both very fast and with high precision. This process is driven by the dynamic changes in signalling activity of the conserved Hedgehog (Hh) pathway, which coordinates cell fate determination, cell cycle and tissue morphogenesis. Here we show that during Drosophila retinogenesis, the retinal determination gene dachshund (dac) is not only a target of the Hh signaling pathway, but is also a modulator of its activity. Using developmental genetics techniques, we demonstrate that dac enhances Hh signaling by promoting the accumulation of the Gli transcription factor Cubitus interruptus (Ci) parallel to or downstream of fused. In the absence of dac, all Hh-mediated events associated to the morphogenetic furrow are delayed. One of the consequences is that, posterior to the furrow, dac- cells cannot activate a Roadkill-Cullin3 negative feedback loop that attenuates Hh signaling and which is necessary for retinal cells to continue normal differentiation. Therefore, dac is part of an essential positive feedback loop in the Hh pathway, guaranteeing the speed and the accuracy of Drosophila retinogenesis.
2016-07-01 | GSE82151 | GEO
Project description:Paracrine Hedgehog (Hh) signaling regulates growth and patterning in many Drosophila organs. We mapped chromatin binding sites for Cubitus interruptus (Ci), the transcription factor that mediates outputs of Hh signal transduction, and we analyzed transcription profiles of control and mutant embryos to identify genes that are regulated by Hh. Putative targets we identified include several Hh pathway components, most previously identified targets, and many targets that are novel. Analysis of expression patterns of pathway components and target genes gave evidence of autocrine Hh signaling in the optic primordium of the embryo. And, every Hh target we analyzed that is not a pathway component appeared to be regulated by Hh in a tissue-specific manner. We present evidence that Hh-dependent tissue specificity is dependent upon transcription factors that are Hh-independent, suggesting that “pre-patterns” of transcription factors partner with Ci to make Hh-dependent gene expression position-specific. Analysis of the expression profiles of loss of function mutantations in core components of the Hh signaling pathway. A total of 14 samples were analysed consisting of comparisons of hh-, ci-, smo-, ptc-, and Cim1-m4 (Activator) mis-expression embryos compared to wt sibling embryos.
2010-11-03 | E-GEOD-24028 | biostudies-arrayexpress
Project description:Aberrant activation of Hedgehog (HH) signaling has been identified as a key etiologic factor of many human malignancies. Signal strength, target gene specificity, and oncogenic activity of HH signaling profoundly depend on interactions with other pathways such as epidermal growth factor receptor-mediated signaling which has been shown to cooperate with HH/GLI in basal cell carcinoma and pancreatic cancer. We demonstrate that the human medulloblastoma cell line Daoy possesses a fully inducible endogenous HH pathway. Treatment of Daoy cells with Sonic Hedgehog or Smoothened agonist induced expression of GLI1 protein and prevented processing of GLI3 to its repressor form. To study interactions between HH- and EGF-induced signaling in greater detail, time-resolved measurements were carried out and analyzed on the transcriptomic as well as proteomic level. Daoy cells responded to the co-treatment by downregulating GLI1, PTCH, and HHIP on the transcript level which was also seen when Amphiregulin (AREG) was used instead of EGF. The finding that EGFR signaling silences proteins acting as negative regulators of HH signaling is firstly described here as a novel crosstalk mechanism. Furthermore, combined EGFR/HH signaling maintains high GLI1 protein levels contrasting its downregulation on the transcript level. On the other hand, high level synergism was observed with respect to a strong and significant upregulation of numerous canonical EGF-targets with putative tumor-promoting properties such as MMP7, VEGFA, and IL-8. In conclusion, synergistic effects between EGFR and HH signaling can selectively induce a switch from a canonical HH/GLI profile to a modulated specific target gene profile pointing to more wide-spread, yet context-dependent, interactions between HH/GLI and growth factor receptor signaling in human malignancies. To study interactions between HH- and EGF-induced signaling, time-resolved measurements were carried out over a period of 24 h at 14 different time points after stimulation by EGF with and without additional stimulation by SHH. Furthermore, as a control, cells without any stimulation by EGF and SHH (control) and cells in the presents of SHH were analyzed. Overall, three biological replicates of 60 different treatment/timepoints were analyzed yielding 180 different samples.
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2013-05-21 | GSE47113 | GEO
Project description:We report the changes in chromatin accessibility associated with regeneration in wing imaginal discs at two developmental timepoints during L3. Wing discs were ablated using tissue-specific expression of the pro-apoptotic gene eiger at an early timepoint (day 7) when the discs retain high regenerative capacity and at a late time point (day 9) when regenerative capacity has significantly declined. ATAC-seq was performed on these discs after 40 hr of ablation and compared to identically staged unablated discs. We detect 14,142 open chromatin peaks after merging overlapping peaks from 3 biological repeats of all 4 conditions. Of these, 349 change significantly (log2fc>0.5, p<0.1) either opening or closing upon damage in early L3 discs, while only 55 open or close in late L3 discs. We describe two consequential groups of peak changes, those that are damage responsive (DR) in early L3 (222 peaks) and those that are maturity silenced (MS), open in early damaged L3 and closed in late damaged L3 (729 peaks). 28 peaks fall in the overlap of these two groups.
2020-06-11 | GSE140755 | GEO