Expression data from Drosophila S2* cells treated with the steroid hormone 20-hydroxy-ecdysone (20E)
Ontology highlight
ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE46020 | GEO | 2013/04/13
SECONDARY ACCESSION(S): PRJNA196913
REPOSITORIES: GEO
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Project description:Previously we and other teams have found that 20E modulates the induction and expression of antimicrobial peptides (AMPs) in immune-challenged Drosophila cell culture or whole animals. To identify potential downstream targets of 20E involved in modulating the immune response we used Affymetrix gene chips (microarrays) to compare the transcriptomes of Drosophila S2* cells treated or not with 20E for 24 hours. Affymetrix Drosophila 2.0 Chips were probed in triplicate with RNA isolated from untreated S2* cells or cells treated with 1 microMolar 20E (Sigma) for 24 hours. cDNA products were hybridized at the Brown University Genomics Core Facility to Affymetrix GeneChip Drosophila_2.0 Genome Arrays (3 replicate chips per treatment).
2013-04-13 | E-GEOD-46020 | biostudies-arrayexpress
Project description:Male Anophele gambiae mosquitoes transfer the steroid hormone 20-hydroxyecdysone (20E) to the female lower reproductive tract during mating. We have shown that this sexually transferred hormone is a key regulator of monandry and oviposition in An. gambiae females. In order to identify possible molecular pathways underlying the effects induced by sexually transferred 20E we investigated the transcriptional response in the atrium and spermatheca at 24 hours after 20E injection in the thorax of virgin females, which delivers to these reproductive tissues 20E levels comparable to those detected after mating. By comparing these results to those of microarrays assessing the post-mating transcriptional response in the female lower reproductive tract (LRT) at 24 hours post copulation, we were able to identify mating responsive genes likely regulated by male transferred 20E. For each experiment atrium and spermatheca were dissected from females 24 hours after 20E or ethanol (10%) injection and transcriptional profiles compared across 4 biological replicates using Agilent two-color microarrays
2015-02-03 | E-MTAB-3023 | biostudies-arrayexpress
Project description:To examine the effects of 20E treatment on mosquito gene expression, treated mosquito cells were examined by RNA-seq to determine the influence of 20E treatment.
2020-04-22 | GSE116252 | GEO
Project description:To identify 20E-regulated genes, wandering third instar larvae were dissected and their organs were cultured in the presence of either no hormone, 20E alone, cycloheximide alone, or 20E plus cycloheximide for six hours. Keywords: Hormone response
2005-10-04 | GSE3060 | GEO
Project description:Juvenile hormone (JH) and 20-hydroxy-ecdysone (20E) are highly versatile hormones, coordinating development, growth, and reproduction in insects. Pulses of 20E provide key signals for initiating developmental and physiological transitions, while JH promotes or inhibits these signals in a stage-specific manner. Previous evidence suggests that JH and 20E might modulate innate immunity, but whether and how these hormones interact to regulate the immune response remains unclear. Here we show that JH and 20E have antagonistic effects on the expression of antimicrobial peptides (AMPs) in Drosophila melanogaster. In S2* cells challenged with bacterial peptidoglycans, 20E induces promoter activity and expression of AMPs in a dose-dependent manner, while JH III and its synthetic analogs (JHa) methoprene and pyriproxyfen abolish this 20E-dependent response. Using microarrays and GFP reporter gene assays in adult flies, we confirm that JH is a hormonal immuno-suppressor in vivo. When silencing both partners of the ecdysone receptor (EcR ) / ultraspiracle (USP) heterodimer with RNAi in S2* cells, 20E fails to activate Diptericin (Dpt) expression, suggesting that 20E regulates expression of this gene through EcR / USP signaling. In contrast, silencing methoprene-tolerant (MET), a candidate JH receptor, does not impair the immuno-suppressive action of JH III and JHa, indicating that in this context MET does not function as a JH receptor. Our results suggest that the balance of 20E and JH is a major determinant of immune homeostasis in insects. Keywords: Topical hormone treatment of whole flies
2007-12-31 | GSE9001 | GEO
Project description:To identify 20E-regulated genes, wandering third instar larvae were dissected and their organs were cultured in the presence of either no hormone, 20E alone, cycloheximide alone, or 20E plus cycloheximide for six hours. Experiment Overall Design: Eight partial blue gut third instar larvae were dissected in each well of a 9 well glass dish (Corning) and cultured in ~100 µl oxygenated Schneiders Drosophila Medium (Gibco) at 25°C. Cultures were incubated in an a styrofoam box under a constant flow of oxygen. Following an initial incubation of 1 hour, the medium was removed and replaced with either fresh Schneiders Drosophila Medium (no hormone), medium plus 8.5x10-5 M cycloheximide (Sigma), medium plus 5x10-6 M 20-hydroxyecdysone (Sigma), or medium plus cycloheximide and 20E, each for 6 hrs at 25°C. Organs were collected and RNA extracted from these samples was analyzed on Affymetrix Drosophila Genome Arrays.
2007-10-31 | E-GEOD-3060 | biostudies-arrayexpress
Project description:Juvenile hormone (JH) and 20-hydroxy-ecdysone (20E) are highly versatile hormones, coordinating development, growth, and reproduction in insects. Pulses of 20E provide key signals for initiating developmental and physiological transitions, while JH promotes or inhibits these signals in a stage-specific manner. Previous evidence suggests that JH and 20E might modulate innate immunity, but whether and how these hormones interact to regulate the immune response remains unclear. Here we show that JH and 20E have antagonistic effects on the expression of antimicrobial peptides (AMPs) in Drosophila melanogaster. In S2* cells challenged with bacterial peptidoglycans, 20E induces promoter activity and expression of AMPs in a dose-dependent manner, while JH III and its synthetic analogs (JHa) methoprene and pyriproxyfen abolish this 20E-dependent response. Using microarrays and GFP reporter gene assays in adult flies, we confirm that JH is a hormonal immuno-suppressor in vivo. When silencing both partners of the ecdysone receptor (EcR ) / ultraspiracle (USP) heterodimer with RNAi in S2* cells, 20E fails to activate Diptericin (Dpt) expression, suggesting that 20E regulates expression of this gene through EcR / USP signaling. In contrast, silencing methoprene-tolerant (MET), a candidate JH receptor, does not impair the immuno-suppressive action of JH III and JHa, indicating that in this context MET does not function as a JH receptor. Our results suggest that the balance of 20E and JH is a major determinant of immune homeostasis in insects. Experiment Overall Design: To examine the transcriptional response of y, w flies to treatment with exogenous JH, we performed a microarray experiment on females treated with JH or solvent (control). Flies were grown on regular yeast diet, switched to no-yeast food within one hour of eclosion, and yeast-starved for 5 days posteclosion to lower their endogenous JH titer and to synchronize their physiology (cf. Tu and Tatar, 2003; Gershman et al., 2007). Subsequently, we anesthetized flies on ice and topically treated them with 0.1 ïl of 187 mM JH III in acetone or with 0.1ï ïl 100% acetone (control) using a 1 ïl Hamilton syringe with a repeating dispenser. 12 hours after hormone administration, samples were snap-frozen in liquid nitrogen and stored at -80°C. RNA was isolated from samples (2 JH samples, 2 control samples, each with 30 females) by lysis, as described in Gershman et al. (2007). cDNA products were hybridized at the Brown University Genomics Core Facility to Affymetrix GeneChip Drosophila_1 Genome Arrays (2 replicate chips per treatment). The dataset consisted of 14,009 probe sets, with 6,142 probe sets annotated. Expression data were analyzed for significant over- or underrepresentation of gene ontology (GO) terms with the web application FatiGO (Al-Shahrour et al., 2004), using a two-fold change criterion. To test whether JH treatment significantly suppresses expression of AMPs we used t-tests implemented in JMP IN 5.1.
2008-06-16 | E-GEOD-9001 | biostudies-arrayexpress
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.
2017-11-01 | GSE92933 | GEO
Project description:The current work describes molecular mechanism used by the Negative elongation complex (NELF) to control the Pol II pause at genes whose transcription is induced by 20-hydroxyecdysone (20E). According to our data NELF complex is recruited to the promoters and enhancers of 20E-dependent genes. Its presence at the regulatory sites of 20E-dependent genes correlates with the described interaction of its NELF-A subunit with EcR receptor. The NELF complex, consisting of at least three subunits, is formed at the 20E-dependent promoters and participates in their active transcription and maintenance of their repressed state. NELF depletion causes a significant decrease in transcription induced by 20E. We associate this effect with the disruption of Pol II elongation complexes formation. A considerable reduction in the promoter-bound level of Spt5 subunit of DSIF was observed at the 20E-dependent genes upon NELF depletion. We presume that participation in stabilization of the Pol II-DSIF complex can be an important function of NELF with a big impact on transcription of its target genes. Part of our study was to directly link NELF to regulation of 20E-dependent genes in development. We showed presence of NELF at the promoters of 20E-dependent genes during their active transcription both in embryogenesis and metamorphosis. We also demonstrate that 20E-dependent promoters remain in a Pol II paused state at the larva stage, where they are temporarily repressed.
2021-01-10 | GSE156847 | GEO
Project description:Insects, unlike vertebrates, are generally believed to lack steroid hormones with functions predominantly associated with adult male biology. In the malaria mosquito Anopheles gambiae, the ecdysteroid 20-hydroxyecdysone (20E) appears to both control egg development in females and induce mating refractoriness and oviposition when sexually transferred by males. Here we show that these sex-specific functions are instead carried out by distinct steroids. We identify a male-specific oxidized form of 20E (3D20E) that upon sexual transfer switches off female mating receptivity, ensuring male paternity. Endogenous female 20E does not induce mating refractoriness, while it triggers oviposition in mated females when expression of a 20E-inhibiting kinase is repressed. 3D20E and 20E have different downstream targets, with 3D20E inducing expression of a tolerance factor that preserves female fitness during Plasmodium infection. The evolution of this male steroid has therefore not only shaped the mating biology of An. gambiae, but also impacted malaria transmission.
2022-04-21 | PXD032157 | Pride