Drosophila Embryos: Control Wild Type vs. Engrailed GooseberryNeuro double heterozygous
Ontology highlight
ABSTRACT: Transcriptional profiling of drosophila embryos (0-14h collection) comparing control wild type embryos with embryos double heterozygous for Engrailed and GooseberryNeuro Two conditions experiment, Wild Type vs. Double heterezygous Engrailed GooseberryNeuro. Biological replicates: 3 control, 3 mutant conditions, independently selected. Triplicate per array
Project description:Transcriptional profiling of drosophila embryos (0-14h collection) comparing embryos double heterozygous for Engrailed and GooseberryNeuro with control wild type embryos
Project description:RNA profiling of Drosophila sensory organs precursor cells (SOP/pI) compared with neighbouring epithelial cells. Two-condition experiment, SOP/pI vs. epithelial cells from animals at similar developmental age.
Project description:The abstract of the manuscript titled "Signalling pathways involved in adult heart formation revealed by gene expression profiling in Drosophila" is given below: "Drosophila provides a powerful system for defining the complex genetic programs that drive organogenesis. Under control of the steroid hormone ecdysone, the adult heart in Drosophila forms during metamorphosis by a remodelling of the larval cardiac organ. Here, we evaluated the extent to which transcriptional signatures revealed by genomic approaches can provide new insights into the molecular pathways that underlie heart organogenesis. Whole-genome expression profiling at 8 successive time-points covering adult heart formation revealed a highly dynamic temporal map of gene expression through 13 transcript clusters with distinct expression kinetics. A functional atlas of the transcriptome profile strikingly points to the genomic transcriptional response of the ecdysone cascade, and a sharp regulation of key components belonging to a few evolutionary conserved signalling pathways. A reverse genetic analysis provided evidence that these specific signalling pathways are involved in discrete steps of adult heart formation. In particular, the Wnt signalling pathway is shown to participate in inflow tract and cardiomyocyte differentiation, while activation of the PDGF-VEGF pathway is required for cardiac valve formation. Thus, a detailed temporal map of gene expression can reveal signalling pathways responsible for specific developmental programs and provides here substantial grasp into heart formation." Keywords: Time-course analysis Total RNAs were prepared from dissected cardiac tubes of Drosophila staged pupae at 8 successive time-points 21, 24, 27, 30, 33, 36, 42 and 48 hours after puparium formation covering adult heart organogenesis. For each time-point sample, 5 cardiac tubes were hand dissected from staged individuals. Four independent biological replicates were analyzed to confer a high reproducibility and statistical significance of the expression data. PolyA+ RNAs were linearly amplified (32 amplifications), labelled (32 with Cy3 and 32 with Cy5), and used for hybridization on Drosophila high-density oligonucleotide (INDAC) dual-channel microarrays (32 arrays) according to a loop-design dedicated to time-course experiments. Given the same number of arrays (8), a simple loop design is more efficient than a reference design: direct comparisons, smallest variance for log ratios, balancing varieties with dye-swapping (including technical replicates) and no reference sample needed (which has no intrinsic interest in our study). A graphical representation of the loop design used on this study is showed in the associated manuscript.
Project description:Engrailed homeoproteins are expressed in adult dopaminergic neurons of the substantia nigra. In Engrailed1 heterozygous mice, these neurons start dying at 6 weeks, are more sensitive to oxidative stress and progressively develop traits similar to those observed following an acute and strong oxidative stress inflected to wild-type neurons. These changes include DNA strand breaks and the modification (intensity and distribution) of several nuclear and nucleolar heterochromatin marks. Engrailed1 and Engrailed2 are biochemically equivalent transducing proteins previously used to antagonize dopaminergic neuron death in Engrailed heterozygous mice and in mouse models of Parkinson disease. Accordingly, we show that, following an acute oxidative stress, a single Engrailed2 injection restores all nuclear and nucleolar heterochromatin marks, decreases the number of DNA strand breaks and protects dopaminergic neurons against apoptosis. RNA-seq data for differentially expressed genes in the SNpc of En1+/- mice, En2 infused mice and 6-OHDA/En2 injection experiments.
Project description:Engrailed homeoproteins are expressed in adult dopaminergic neurons of the substantia nigra. In Engrailed1 heterozygous mice, these neurons start dying at 6 weeks, are more sensitive to oxidative stress and progressively develop traits similar to those observed following an acute and strong oxidative stress inflected to wild-type neurons. These changes include DNA strand breaks and the modification (intensity and distribution) of several nuclear and nucleolar heterochromatin marks. Engrailed1 and Engrailed2 are biochemically equivalent transducing proteins previously used to antagonize dopaminergic neuron death in Engrailed heterozygous mice and in mouse models of Parkinson disease. Accordingly, we show that, following an acute oxidative stress, a single Engrailed2 injection restores all nuclear and nucleolar heterochromatin marks, decreases the number of DNA strand breaks and protects dopaminergic neurons against apoptosis.
Project description:A number of homeobox genes have been found to be implicated in the development of various cancers. Here, we investigated the role of engrailed 2 (EN2), a member of the homeobox gene superfamily, in esophageal squamous cell carcinoma
Project description:Genome-wide binding profile of PcG proteins Pho and Ph, H3K27me3 and Engrailed in Drosophila third instar larval brains and disks, in duplicate, using Illumina HiSeq 2500. The sequence reads that passed quality filters were analyzed. Drosophila third instar larval brains and disks were dissected and fixed with 2% HCHO before sonication. After sonication, chromatin immunoprecipitation using anti-Pho, -Ph, -H3K27me3, and -Engrailed antibodies was carried out.
Project description:Transcriptional profiling of the preingression epiblast versus lateral epiblast, of preingression epiblast from control embryos treated with the DMSO carrier vehicle (0.5%) versus embryos treated with SU5402, U0126 or LY294002. Embryos were stages 5-6 at time of tissue isolation. The preingression epiblast (E2 region) is the region of epiblast just lateral to the primitive streak. Lateral epiblast is the E3 region. Two condition experiment. Samples for each condition isolated from at least 30 embryos. The experiments used isolated preingression epiblast, which is defined as the region of epiblast adjacent to the primitive streak that will ingress through the primitive streak. Preingression epiblast was isolated by microdissection using tungsten needles from control embryos or embryos treated with SU5402, U0126 or LY294002.