Project description:This SuperSeries is composed of the following subset Series: GSE11311: Drosophila Sex Hierarchy Regulated Gene Expression in 48 hour APF Pupae GSE11313: Drosophila Gene Expression During Metamorphosis in Wild Type and Germline Minus Pupae Keywords: SuperSeries Refer to individual Series
Project description:We used long-oligonucleotide microarrays to investigate whether alternative splicing in Drosophila is regulated in a sex-, stage-, or tissue-specific manner. To examine sex-specific splicing, we compared gene expression profiles of male and female pupae 12 hours after pupariation. To examine stage-specific splicing, we compared expression profiles of mixed-sex, 0-24 hour old embryos and mixed-sex, 12 hour old pupae. To examine tissue-specific splicing, we compared expression profiles of adult male heads and abdomens 24-48 hours after eclosion. To examine tissue-specific splicing, we compared expression profiles of adult male heads and abdomens at 24-48 hours after eclosion. Keywords: tissue-specific expression profiles
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.
Project description:We used long-oligonucleotide microarrays to investigate whether alternative splicing in Drosophila is regulated in a sex-, stage-, or tissue-specific manner. To examine sex-specific splicing, we compared gene expression profiles of male and female pupae 12 hours after pupariation. To examine stage-specific splicing, we compared expression profiles of mixed-sex, 0-24 hour old embryos and mixed-sex, 12 hour old pupae. To examine tissue-specific splicing, we compared expression profiles of adult male heads and abdomens 24-48 hours after eclosion. To examine tissue-specific splicing, we compared expression profiles of adult male heads and abdomens at 24-48 hours after eclosion. Keywords: tissue-specific expression profiles Drosophila isogenic line WI89 was used. Mixed-sex, mixed-stage embryos were harvested from plates on which females had been allowed to oviposit for 24 hours. To obtain synchronized cohorts of pupae, male and female white prepupae were collected at 0-1 hour after pupariation and aged for 12 hours at 25C. Mixed-sex pupal samples were generated by mixing equal amount of male and female pupal RNA. Adult heads and abdomens were dissected from 24-48 hour old males. mRNA was isolated and labeled without amplification.
Project description:In Drosophila, male-specific FRU (FRUM) is required to establish the potential for courtship behaviors, but the downstream effectors of FRUM during development are largely unknown. A microarray-based approach identified genes that are differentially expressed as a consequence of FRUM in pupae, in both whole body and CNS tissues. Genes were also identified that are sex-differentially expressed in CNS tissues. The FRUM-regulated sets were significantly overrepresented with genes also regulated by the ecdysone regulatory pathway. Two EcR isoforms (EcRA and EcRB1) are expressed in FRUM-expressing neurons during distinct periods of metamorphosis. Males with abrogated EcRA function in FRUM-expressing neurons aggressively court other males. Transcriptional profiles of mutants with abrogated EcRA function in the fru circuit demonstrate that EcR and FRUM regulate common gene sets, including the early gene broad. These results demonstrate a novel role for EcR in specifying male courtship behavior through its actions specifically in the FRUM neural circuitry. All microarrays were dual channel with direct comparisons of male versus female, wild type versus mutant, or experimental versus control. For each experiment, four to eight independent biological samples were analyzed using a dye-swap design. Samples consisted of whole body pupae or dissected CNS collected either at 0 hour After Pupal Formation (APF), 48 hour APF, or 0-24 hour adults.
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
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.
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).