Differentially expressed genes at molting and metamorphosis stages in Bombyx fat body
Ontology highlight
ABSTRACT:
ORGANISM(S): Bombyx mori
PROVIDER: GSE23424 | GEO | 2010/08/05
SECONDARY ACCESSION(S): PRJNA131043
REPOSITORIES: GEO
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Project description:In the silkworm, Bombyx mori, juvenile hormone (JH) and 20-hydroxyecdysone (20E) levels are high during the final larval molt (4M) but both absent during the feeding stage of 5th instar (5F), while JH level is low and 20E level is high during the prepupal stage (PP). Fat body is the important organs in insect, we want to find out differentially expressed genes which are respectively regulated by the two hormones. Total RNA from 4th molting,5th feeding and prepupa stages Bombyx fat body were used to generate target cDNA, and then hybridized to 48k Bombyx genome Array Genechips, representing about 23000 characterized genes
2010-08-04 | E-GEOD-23424 | 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. Keywords: Topical hormone treatment of whole flies
2007-12-31 | GSE9001 | 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. 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.
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Project description:This study identifies genes that alter their expression in synchrony with the late third instar and prepupal pulses of 20E. Keywords: time course
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Project description:Juvenile hormone (JH) is produced and secreted by corpora allata (CA) during larval stages. JH biosynthesis from acetyl-CoA includes 13 enzymatic steps. Genes for 10 of those enzymes have been identified from Bombyx mori. It was also reported that they are basically expressed selectively in CA. Genes for the other 3 enzymes were recently reported from other insects, i.e., farnesyl diphosphate pyrophosphatase (FPPP) from Drosophila and farnesol dehydrogenase (FolD) and farnesal dehydrogenase (FalD) from Aedes. In order to reveal the expression profiles of these genes in Bombyx CA and to find other candidate genes involved in JH biosynthesis, we performed a transcriptomic analysis in corpora cardiaca-CA (CC-CA) complexes using a custom-made DNA microarray on which sequences from CC-CA EST libraries and genome-widely predicted genes are loaded. CC-CA complexes vigorously expressed JH-biosynthetic genes identified in Bombyx as well as genes involved in methyl-group metabolism and ribosomal protein genes; this is suitable for JH production. Bombyx counterparts of Drosophila FPPP and Aedes FolD genes, however, showed quite low level of expression. On the other hand, some other genes probably encoding oxidoreductases were expressed highly and selectively in CC-CA complex. Their products might function as FolD and/or FalD in Bombyx.
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Project description:Juvenile hormone (JH) antagonizes 20-hydroxyecdysone (20E) signaling mainly through the intracellular receptors Met and Gce in Drosophila. To investigate JH membrane signaling pathway without the interference from JH intracellular signaling, we characterized phosphoproteome profiles of the Met gce double mutant in the absence or presence of JH in both chronic and acute phases. Importantly, JH was identified to phosphorylate USP at Ser35, the PKC phosphorylation site required for the maximal action of 20E through its nuclear receptor complex EcR-USP. Functioning through a potential receptor tyrosine kinase and phospholipase C pathway, JH membrane signaling activated PKC to phosphorylate USP at Ser35. The USPS35A mutant, in which Ser35 was replaced with Ala35 by genome editing, showed attenuated 20E signaling and Yorkie activity, resulting in delayed developmental timing and reduced body size, respectively. Moreover, genetic interaction experiments confirmed that JH lied upstream of PKC-mediated USP phosphorylation at Ser35. This study propose a novel model of JH membrane signaling that exhibits crosstalk with different pathways simultaneously to regulate distinct developmental processes.
2020-09-23 | PXD021644 |
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
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:This study identifies genes that alter their expression in synchrony with the late third instar and prepupal pulses of 20E. Experiment Overall Design: RNA was isolated from w1118 whole animals staged at -18, -4, 0, 2, 4, 6, 8, 10, and 12 hours relative to pupariation, labeled, and hybridized to Affymetrix Drosophila Genome Arrays.
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Project description:Variations of hepatopancreatic transcript expression related to induced molt cycle were identified in male Cherax quadricarinatus using a cDNA microarray estimated to contain 2180 unique sequences. Molt induction was performed by X-organ sinus gland extirpation or by repeated 20-hydroxyecdysone injections. Manipulated males were sacrificed at premolt or early postmolt and a reference population at intermolt. Their isolated hepatopancreatic mRNA was hybridized onto the microarray and differentially-expressed genes were identified, sequenced, and annotated. Clusters of similarly expressed transcripts, in correlation with the four combinations of induction methods and molt stages were identified, containing both known and novel genes. Biologically interesting clusters were characterized by general shift of expression throughout premolt and early postmolt vs. intermolt, or by different premolt vs. postmolt expression. Several genes were differentially expressed in 20-hydroxyecdysone injected crayfish vs. X-organ sinus gland extirpated ones. Keywords: Physiological state analysis
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