Drosophila Sex Hierarchy Regulated Gene Expression in 48 hour APF Pupae
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ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE11311 | GEO | 2009/02/11
SECONDARY ACCESSION(S): PRJNA109201
REPOSITORIES: GEO
Similar Datasets
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
Project description:The Drosophila sex determination hierarchy controls all aspects of somatic sexual differentiation, including sex-specific differences in adult morphology and behavior. To gain insight into the molecular-genetic specification of reproductive behaviors and physiology, we identified genes expressed in the adult head and central nervous system that are regulated downstream of sex-specific transcription factors encoded by doublesex (dsx) and fruitless (fru). We used a microarray approach and identified 54 genes regulated downstream of dsx. Furthermore, based on these expression studies we identified new modes of DSX-regulated gene expression. We also identified 90 and 26 genes regulated in the adult head and central nervous system tissues, respectively, downstream of the sex-specific transcription factors encoded by fru. In addition, we present molecular-genetic analyses of two genes identified in our studies, calphotin (cpn) and defective probocisis response (dpr), and begin to describe their functional roles in male behaviors. We show that dpr and dpr-expressing cells are required for the proper timing of male courtship behaviors. Keywords: genetic modification
2007-11-06 | GSE9185 | 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: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. Genes were identified that were expressed during metamorphosis in both somatic and germline tissues of males and females. Additionally, genes were identified that display sex-specific differences in abundance in both of these tissues at discrete times during metamorphosis. Keywords: time course; wild type; genetic modification; Gene expression was examined at five time points during metamorphosis: 0, 24, 48, 71, and 96 hr After Puparium Formation (APF). Gene expression was examined separately in males and females for both wild type pupae and tudor (tud) progeny. tud progeny have genetically ablated germline tissues. All samples were labeled with Cy5 and compared against a common reference sample labeled with Cy3. The reference sample contained male and female wild type pupae from all stages of metamorphosis. All experiments were conducted in triplicate.
2009-02-10 | E-GEOD-11313 | biostudies-arrayexpress
Project description:Purpose: Sexual dimorphisms fuel significant intraspecific variation and evolutionary diversification. Yet the developmental-genetic mechanisms underlying sex-specific development remain poorly understood. We focused on the conserved sex-determination gene doublesex (dsx) and the mechanisms by which it mediates sex-specific development in a species of horned beetle by combining systemic dsx knockdown, high-throughput sequencing of diverse tissues, and a genome-wide analysis of Dsx binding sites. We find that Dsx regulates sex-biased expression predominantly in males, that Dsx's target repertoires are highly sex- and tissue-specific, and that Dsx can exercise its regulatory role via two distinct mechanisms: as a sex-specific modulator by regulating strictly sex-specific targets, or as a switch by regulating the same genes in males and females in opposite directions. More generally, our results suggest Dsx can rapidly acquire new target gene repertoires to accommodate evolutionarily novel traits, evidenced by the large and unique repertoire identified in head horns, a recent morphological innovation.
2017-02-28 | GSE87788 | GEO
Project description:We recently identified a missense mutation in Nucleoporin107 (Nup107; D447N) underlying XX-ovarian-dysgenesis (XX-OD), a disorder characterized by underdeveloped and dysfunctional ovaries. Specific knockdown of Nup107 in the somatic gonadal cells and moreover, modelling of the human mutation in Drosophila result in ovarian-dysgenesis-like phenotypes in female flies. The aberrant phenotypes in larval and adult ovaries compromised for Nup107 are associated with hyperactivation of BMP signalling. Transcriptomic analysis identified the somatic sex-determination gene Doublesex (dsx) and the extracellular metalloprotease AdamTS-A as targets of Nup107. Either loss or gain of Dsx in the gonadal soma is sufficient to respectively mimic or rescue the phenotypes induced by Nup107 loss. Furthermore, adamTS-A is transcriptionally regulated by Dsx, and its knockdown in the somatic gonad hyperactivates BMP signaling and to a large extent recapitulates loss of Nup107 phenotypes. Thus, Dsx acts downstream of Nup107 to impact female germline stem cells via sex-specific modulation of the BMP pathway.
2022-03-21 | GSE141094 | GEO
Project description:Investigating Drosophila melanogaster genome-wide gene expression differences in developing pupal eyes comparing 21 hours After puparium formation (APF) to 40 hours APF.
2020-10-30 | GSE160441 | GEO
Project description:The Drosophila sex determination hierarchy consists of a splicing cascade with sex-specific transcription directing somatic sexual dimorphism. Our understanding of this pathway, and many others, is incomplete. Here we pioneer an approach to expand our knowledge of gene regulatory networks (GRNs) by leveraging natural genetic variation. This approach is generalizable to any natural population, including humans. Two studies from Drosophila female head tissue were used – the DSPR collection (alleles from 15 natural variants) and F1-hybrid collection (alleles from heterozygotes of 75 isogenic lines crossed to w1118) – in a structural equation model (SEM) analysis. We expanded the sex hierarchy GRN by adding novel links among genes in the pathway and by adding novel genes to the pathway. A link from fruitless (fru) to Sex-lethal (Sxl) was found in both populations, which is supported by the presence of fru binding sites in the Sxl locus. The splicing factors male-specific lethal 2 and Rm62 were correctly identified as downstream targets of Sxl. There were 754 additional candidate genes for an expanded sex hierarchy GRN. These candidates were enriched for genes with sex-biased splicing and many components of the spliceosome were placed in the GRN. As with other population-genetic analyses, the number of alleles limits the number of observable interactions. Network expansion was only clear in the F1-hybrid population, which has an average of twice as many alleles as the DSPR population. Independent studies of doublesex and transformer mutants support many novel connections, including evidence for a link between the sex hierarchy and metabolism, with the inclusion of Insulin-like receptor in the sex hierarchy GRN. RNA sequencing was performed on mRNA derived from adult male or female heads, for a total of 9 samples. These samples included females that produce the male isoform of dsx [w/+;DsxD/dsxm+r15 (XX)], and two dsx mutants: females [w/+; dsxm+r15/dsxd+r3 (XX)] and males [w;dsxm+r15/dsxd+r3 (XY)]. Two wild type genotypes (Berlin and Canton-S) were sequenced at the same time, but have previously been published as part of GSE50515. There were at least 3 replicates from biological samples.
2015-09-01 | E-GEOD-67400 | biostudies-arrayexpress
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.
2011-02-25 | GSE23344 | GEO
Project description:The survival and reproductive success of animals depends on the ability to harmonize their external behaviors with their internal states. For example, females conduct numerous social programs that are distinctive to virgins compared to post-mated and/or pregnant individuals. In Drosophila, the fact that the post-mating switch is initiated by seminal factors implies the default state as virgin. Recently, we uncovered molecular and genetic control of the female virgin state. In particular, repression of the transcription factor Homothorax (Hth) by mir-iab-4/8 within the abdominal ventral nerve cord (VNC) is critical for unmated females to execute virgin behaviors. To elucidate new components of this regulatory circuit, we exploited mir-iab-4/8 deletion and hth-miRNA binding site mutants (hth[BSmut]) to elucidate doublesex (dsx) as a critical downstream factor. While Dsx has mostly been studied during sex-specific differentiation, its activities in neurons are little known. We find that accumulation of Dsx in the CNS is mutually exclusive with Hth, and downregulated in miRNA/hth[BSmut] mutants. Moreover, experimental suppression of Dsx in highly restricted sets of abdominal neurons abrogates female virgin conducts, in favor of mated behavioral programs. Thus, a double negative pathway in the VNC mediates the virgin behavioral state.
2021-06-29 | GSE166562 | GEO