An atlas of transcription factors expressed in the Drosophila melanogaster pupal terminalia
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ABSTRACT: The male terminalia of Drosophila melanogaster has undergone rapid morphological change between it and its closely related species, making it a useful model for studying evolution and development. In order to identify the network components that pattern the male terminalia, we isolated and sequenced whole RNA from two timepoints during pupal development, and next performed in situ hybridization for 100 highly expressed transcription factors during pupal development.
Project description:The goal of this gene expression profiling experiment was to identify the entire set of transcription factors expressed during late pupal wing development (~80h APF) when pigmentation genes are expressed
Project description:The goal of this gene expression profiling experiment was to identify the entire set of transcription factors expressed during late pupal wing development (~80h APF) when pigmentation genes are expressed We used Affymetrix microarrays to profile the expression of the transcription factors in late pupal wings of Oregon R flies. The staging of the flies was made by eye, based on the extent of wing pigmentation. RNA was extracted from batches of 40 flies and three biological replicates were analyzed.
Project description:To examine the repertoires of genes expressed in individual fru P1-expressing neurons, we performed single cell sequencing. The analysis was performed on male and female central nervous system tissues (48-hour pupal stage), from flies that expressed membrane-bound GFP in fru P1 neurons. We chose this developmental stage to gain further insight into how genes direct development of fru P1 neurons, as this is the stage where FruM has peak expression (in ~2,000 neurons).
Project description:We exploit the predictable time course of Drosophila brain development to perform a temporally coupled quantitative proteomic analysis of the pupal brain in Nab2 mutant or overexpression models, which reveals that Nab2 is required to regulate the abundance of a number of proteins with critical roles in Drosophila neurons. Pupal brains lacking Nab2 show dysregulation of proteins, such as Futsch, Turtle, Contactin, and Van Gogh, that typically function in brain morphogenesis, neuroblast proliferation, circadian sleep/wake cycle, and other neurodevelopmental processes. Overall, these data define a role for Nab2 during neurodevelopment in regulating protein abundance for a subset of the brain proteome and provide a window into the potential functions of human ZC3H14 protein.
Project description:Custom microarrays were used to examine global differences in female vs. male gene expression in the developing pupal head of the dengue vector mosquito Aedes aegypti.
Project description:During development, transcription factors and signaling molecules govern gene regulatory networks to direct the formation of unique morphologies. As changes in gene regulatory networks are often implicated in morphological evolution, mapping transcription factor landscapes is important, especially in tissues that undergo rapid evolutionary change. The terminalia (genital and anal structures) of Drosophila melanogaster and its close relatives exhibit dramatic changes in morphology between species. While previous studies have identified network components important for patterning the larval genital disc, the networks governing adult structures during pupal development have remained uncharted. Here, we performed RNA-seq in whole Drosophila melanogaster male terminalia followed by in situ hybridization for 100 highly expressed transcription factors during pupal development. We find that the male terminalia are highly patterned during pupal stages and that specific transcription factors mark separate structures and substructures. Our results are housed online in a searchable database (https://flyterminalia.pitt.edu/) as a resource for the community. This work lays a foundation for future investigations into the gene regulatory networks governing the development and evolution of Drosophila terminalia.
Project description:Transcriptional regulation by Store-operated Calcium Entry (SOCE) is well studied in non-excitable cells. However, the role of SOCE has been poorly documented in neuronal cells with more complicated calcium dynamics. Previous reports demonstrated a requirement of neuronal SOCE for Drosophila flight. We identified the early pupal stage to be critical and used RNA-sequencing to identify SOCE mediated gene expression changes in the developing Drosophila pupal nervous system. We down-regulated dStim, the endoplasmic reticular calcium sensor and a principal component of SOCE in the nervous system for a 24h period during pupal development, and compared wild type and knockdown transcriptional profiles, immediately after knockdown as well as after a 36h recovery period. We found that dStim knockdown altered the expression of a number of genes. We also characterized one of the down-regulated genes, Ral for its role in flight. Thus, we identify neuronal SOCE as a mechanism that regulates expression of a number of genes during the development of the pupal nervous system. These genes can be further studied in the context of pupal nervous system development.
Project description:Female adult spongy moths vary in their flight capacity depending upon their origin and subspecies. Asian spongy moth (Lymantria dispar asiatica) females (such as the RM strain here) can fly whereas European spongy moth (Lymantria dispar dispar) females cannot (such as the CT strain here). We hypothesized that the genetic origins of these differences in flight capacity would be reflected in differences in gene expression during metamorphosis (the pupal stage). To test this, we compared the gene expression at days 1, 3, 5, 8, and 11 of pupal development of female pupae from CT and RM strains.
Project description:Custom microarrays were used to examine global differences in female vs. male gene expression in the developing pupal head of the dengue vector mosquito Aedes aegypti. RNA was extracted from the heads of male and female 24 hr pupae. 20 male or female heads were pooled for each of four replicates. Hybridization experiments were performed on the Nimblegen Aedes aegypti 12-plex microarray design: 090305_Aedes_aegypti_TEfam_expr.ndf. Four unique replicates and two repeat replicates were assessed in the hybridization experiment.