Project description:Transcriptional analysis of the pheromone gland in comparison to the rest of the insect body in female Agrotis segetum (Noctuidae).
Project description:The tobacco hornworm, Manduca sexta, is a lepidopteran model species widely used to study insect biochemical processes. While some of its larval hemolymph proteins are well understood, a detailed proteomic analysis was unavailable until 2016, revealing features such as correlation with transcriptome data, formation of immune complexes, and constitution of an immune signaling system. Yet, it is unclear how these may change in other developmental stages. In this paper, we report the proteomes of cell-free hemolymph from prepupae, pupae on days 4 and 13, and young adults. Of the 1,824 proteins identified, 907 have a signal peptide and 215 are related to immunity. Drastic changes in abundance of the storage proteins, for instance, reflect physiological disparities among prepupae, pupae, and adults. Considerably more proteins lacking signal peptide are present in the late pupae, suggesting that plasma acts as a temporary reservoir for intracellular components released from remodeling tissues during metamorphosis. In summary, the proteins and their levels revealed in this study are expected to stimulate focused explorations of humoral immunity in wandering larvae, pupae, and adults of M. sexta and shed light upon functional and comparative genomic research in other holometabolous insects.
Project description:Overall, the study aims at obtaining a comprehensive picture of the African malaria mosquito, Anopheles gambiae, transcriptome using high-coverage RNA-seq of sexed whole-insect samples collected at different developmental time points. This experiment focuses on male and female transcriptomes from 20 hour old embryos, 12 hour old 3rd instar larvae, 24 hour old 4th instar larvae and 10 hour old pupae sampled using a strand-specific RNA-seq approach.
Project description:Nilaparvata lugens, or the brown planthopper, is one of the most notorious pest insects of cultured rice, and a model for hemimetabolous development. Recently, the N-glycome of N. lugens was explored throughout post-embryonic development and reproductive stages, which revealed differential protein N-glycosylation events between adult sexes. Identifying the proteins carrying these differential carbohydrate structures would point to the functionality of sex-dependent N-glycosylation. Furthermore, potential effects of the adult wing type on protein N-glycosylation are of interest. Here, a comprehensive investigation of the N-glycosylation sites from the adult stages of N. lugens was conducted, allowing a qualitative and quantitative comparison between sexes and wing forms at the glycopeptide level. N-glycopeptide enrichment using the N-glyco-FASP method with the high mannose/paucimannose-binding lectin Concanavalin A, or the Rhizoctonia solani agglutinin which interacts with complex N-glycans led to the identification of over 1,300 N-glycosylation sites derived from over 600 glycoproteins. Comparison of these N-glycopeptides revealed striking differences in protein N-glycosylation between sexes, while almost no differences were observed between wing types. Male- and female-specific N-glycosylation sites were identified, and some of these sex-specific N-glycosites were shown to be derived from proteins with a putative role in insect reproduction. Transcript expression experiments with complete insects and insect tissues confirmed the sex-related N-glycosylation of proteins, and expression of glycoproteins in reproductive tissues. In conclusion, this study provides original data on N-glycosylation sites of N. lugens adults, providing novel insights into planthopper’s biology and revealing that protein N-glycosylation is sex-related in this insect.
Project description:Expression profiling of honey bee brains exposed to brood pheromone. Exposure was performed in colonies and young (5 days-old) and old bees (15 days-old) were analyzed .
Project description:In Drosophila,male courtship behaviour serves as an excellent paradigm to study how innate behaviors are controlled by the nervous system. These behaviors are in large part regulated by the gene fruitless (fru). fru encodes a set of putative transcription factors that promote male sexual behaior by controlling the development of sexually-dimorphic neuronal circuitry. Little is known about how Fru proteins function at the level of transcriptional regulation. To characterize the roles of fru sex-specific isoforms in specifying male behavior, we generated novel isoform-specific mutants, and used a genomic approach to identify direct Fru isoform targets during development. We demonstrate that all Fru isoforms directly target genes involved in the development of the nervous system, with individual isoforms exhibiting unique binding specificities. We observe that fru behavioral phenotypes are specificed by either a single, or combination, of isoforms. Finally, we illustrate the utility of these data for the identification of novel sexually dimorphic genoic enhancers, and novel downstream regulators of male-sexual behavior. 3 replicates per condition, one dye swap; per condition: one control, one experimental per replicate. No processed data for GSM1261882 and GSM1261883 are available.
Project description:Dosage compensation in D. melanogaster males is achieved via targeting of the MSL complex to X chromosomal genes. This is proposed to involve initial sequence-specific recognition of the X at ~150-300 chromatin entry sites, and subsequent spreading to nearby active genes. Here we test a model in which the spreading step requires transcription and is sequence-independent. We ask whether, in the native context of the X chromosome, MSL complex will target genes of autosomal origin. We find that MSL complex does bind such genes, but only if transcriptionally active. Targeting is accompanied by acetylation of the histone H4K16 residue and two-fold transcriptional up-regulation. We conclude that the presence of a long-sought specific DNA sequence within X-linked genes is not obligatory for MSL complex binding. Instead, physical linkage and transcription play the pivotal roles in the identification of MSL targets irrespective of their origin and DNA sequence. Keywords: Epigenetics ChIP-seq measurements of MSL complex binding and input control. Chromatin was prepared from third instar male larvae of a genotype y w TrojanElephant; MSL3-TAP; msl3. TrojanElephant is a mini-white- and yellow-marked transposition of genomic region spanning 65 kb from cg13773 to snRNP70K. MSL3-TAP is a mini-white-marked TAP-tagged genomic msl3 transgene. IgG beads were used for pull-down.
Project description:Since their discovery, transposable elements have been proposed to play a central role in the evolution of their host genomes through their ability to regulate gene expression, in particular by providing transcription start sites (TSSs) for host genes. To investigate their contribution to developmental gene expression, we developed RAMPAGE, a high-throughput 5'-complete cDNA sequencing approach to accurately discover TSSs, characterize their transcripts, and quantify their expression. This strategy, which directly delineates the expression profiles of individual promoters and was designed to offer optimal sample multiplexing capabilities, represents an advantageous alternative to standard RNA-Seq for a wide range of transcriptome profiling applications. We used RAMPAGE in a genome-wide study of promoter activity throughout 36 stages of the life cycle of Drosophila melanogaster, and describe here a comprehensive dataset that represents the first developmental timecourse of promoter usage. We found that over 40% of developmentally expressed genes have at least 2 promoters, and that alternative promoters generally implement distinct regulatory programs. Transposons harbor TSSs driving the expression of hundreds of annotated genes, and they often impart their own expression specificity upon the genes they regulate. Detailed analysis of particular transposons identified sequence elements encoding these regulatory properties. Our results show that transposable elements contribute significantly to the generation of standing variation and to the evolution of gene regulatory networks, by distributing stereotyped regulatory modules throughout the genome. This dataset represents a whole-genome, single-base resolution profiling of transcription start site (TSS) expression throughout 36 stages of the life cycle of Drosophila melanogaster. These profiles were established using RAMPAGE, a high-throughput, high-accuracy 5'-complete cDNA sequencing method implemented on the Illumina platform. Embryos, larvae, pupae and adult flies were collected at specific stages of development, and RAMPAGE profiles were established for pools of whole organisms. The data was analyzed using custom scripts and algorithms that are all available upon request. Supplementary files: Dmel_Combined_+.bw: bigWig coverage by cDNA 5' ends (+ strand). Dmel_Combined_-.bw: bigWig coverage by cDNA 5' ends (- strand). Dmel_All_RAMPAGE_peaks.bed: BED file describing all RAMPAGE peaks. Dmel_GeneTSS_RAMPAGE_peaks.bed: BED file describing all peaks attributed to annotated genes. GeneTSS_expression_RAMPAGE_RPM.txt: Expression matrix for all genic peaks (RPM: reads per million). Transposon_expression_RAMPAGE_RPM.txt: Expression matrix for all RepeatMasker-annotated transposon classes (RPM: reads per million). Genome build: dm3
Project description:The goal of our microarray experiments was to compare the gene expression profile of two spirodiclofen resistant spider mite strains (SR-VP and SR-TK) with that of a susceptible spider mite strain (LS-VL) 5 samples were analyzed: 3 biological replicates for SR-VP, 2 biological replicates for SR-TK
Project description:We studied the molecular mechanisms underlying the impact of pollen nutrients on honey bee (Apis mellifera) health and how those nutrients improve resistance to parasites. Using digital gene expression, we determined the changes in gene expression induced by pollen intake in worker bees parasitized or not by the mites Varroa destructor, known for suppressing immunity and decreasing lifespan of bees. bees with or without verroa, and fed or not fed pollen