Project description:Using hight throughput sequencing and data processing, we identified miRNA and quantified their expression in the fat body of adult females of Locusta migratoria.
Project description:In honey bees, Vitellogenin (Vg) is hypothesized to be a major factor affecting hormone signaling, food-related behavior, immunity, stress resistance and lifespan. Likewise microRNAs play important roles in posttranscriptional gene regulation and affect many biological processes thereby showing many parallels to Vg functions. The molecular basis of Vg and microRNA interactions is largely unknown. Here, we exploited the well-established RNA interference (RNAi) protocol for Vg knockdown to investigate its effects on microRNA population in honey bee foragerM-bM-^@M-^Ys brain and fat body tissue. To identify microRNAs that are differentially expressed between tissues in control and knockdown foragers, we used M-BM-5ParafloM-BM-. microfluidic oligonucleotide microRNA microarrays. Our results show 76 and 74 miRNAs were expressed in the brain of control and knockdown foragers whereas 66 and 69 miRNAs were expressed in the fat body of control and knockdown foragers respectively. Target prediction identified potential seed matches for differentially expressed subset of microRNAs affected by Vg knockdown. These candidate genes are involved in a broad range of biological processes including insulin signaling, juvenile hormone (JH) and ecdysteroid signaling previously shown to affect foraging behavior. Thus, here we demonstrate a causal link between Vg expression-variation and variation in the abundance of microRNAs in different tissues with possible consequences for regulation of foraging behavior. We knocked down Vitellogenin (Vg) gene expression (using RNAi) in adult workers to identify potential downstream consequences on the expression of microRNA population in the fat body compared to control group (dsRNA-GFP injected bees). Six biological samples of fat body-derived small RNA fraction were prepared for each treatment group (dsRNA-Vg and dsRNA-GFP). Each biological sample contained pooled RNA from 5 unique individuals. Each fat body pool contained a total of 2 M-BM-5g of small RNA fraction, to which each of the 5 individuals contributed equally (400 ng). Pools were named as M-bM-^@M-^\control forager fat bodyM-bM-^@M-^] (GFFb) and M-bM-^@M-^\knockdown forager fat bodyM-bM-^@M-^] (VFFb), followed by a number from 1 to 6.
Project description:Fat body is an important tissue in the context of vitellogenesis, vector immunity, vector physiology and vector-parasite interaction. However, the proteome of this vital organ has not been investigated in any Anopheline species so far. In this study, we employed multiple fractionation method followed by high resolution mass spectrometry to characterize fat body proteome of female mosquitoes An. stephensi Indian strain. In all, we identified 4, 535 proteins in the fat body and a subset of these proteins were found to be restricted to fat body. Gene ontology analysis of these proteins suggested their role in metabolism, lipid transport, vitellogenesis, mosquito immunity and oxidation-reduction processes. By far, this is the largest proteomic resource of fat body in any mosquito species.
Project description:In this work, we aimed to explore miRNA expression and potential targets in the female fat body of the Ae. aegypti mosquito, as well as their changes as a result of blood meal. The fat body is the metabolic center of the insect organism, playing a key role in reproduction. Therefore, understanding of regulatory networks controlling its functions is critical, and the role of miRNAs in the process is largely unknown. Small RNA library analyses revealed four unique miRNA clusters sequentially expressed during the post blood meal (PBM) phase, drawing connections to waves of upstream hormonal signals and gene expression in the same period. To link the miRNA identities with specific downstream targets, transcriptome-wide mRNA 3' UTR interaction sites were experimentally determined at 72 h post eclosion (PE) and 24 h PBM by means of AGO1 CLIP-seq. Hence, the presented manuscript comprehensively elucidated miRNA expression and target dynamics in female mosquito fat body, providing a solid foundation for future functional studies of miRNA regulation during the gonadotrophic cycle.
Project description:Transcript profiling of adult abdominal fat body in S1106>AopACT females after 5 days of induction of transgene expression by RU486 feeding.
Project description:We report the transcriptome profile of one sequenced sample of mRNA isolated from pooled (20 from each genotype) abdomen fly extracts enriched in fat body content of fat body-specific Sdc RNAi knockdown and control flies Abdominal fat body mRNA profiles of 4-6-day old control and fat body-specific Sdc RNAi knockdown were generated by deep sequencing using Illumina HiSeq 2500
Project description:Obesity is a complex disease with many causes, including a possible role of environmental chemicals. Perfluorohexane sulfonate (PFHxS) is one of many per- and polyfluoroalkyl substances (PFASs) frequently detected in humans and it is a suspected obesogenic compound. We examined the potential long-term effects of PFHxS on metabolic parameters in rats after developmental exposure to 0.05, 5 or 25 mg/kg bw/day, with or without co-exposure to a background mixture of 12 endocrine disrupting chemicals (EDmix). Both male and female offspring showed signs of lower birth weight following intrauterine exposure. Female offspring exposed to both PFHxS and EDmix showed increased body weight in adulthood. Furthermore, the retroperitoneal fat pad was larger in these female offspring when compared to those exposed to EDmix alone. An attempt to detect putative molecular markers in the fat tissue by performing whole transcriptome profiling resulted in no significant changes between groups and there were no significant effects on plasma leptin levels in exposed females. These results show that early life exposure to endocrine disrupting chemicals can influence body weight later in life, but the effect is not necessarily reflected in changed gene expression in the fat tissue.
Project description:We performed mRNA-seq from hand-dissected fat body tissue from 68hr (after egg laying, AEL) and 92hr AEL Drosophila melanogaster larvae. Fat body was dissected from wild-type (OrR) males and testes were removed. We examined gene expression genome-wide with particular focus on genes in the underreplicated regions in the fat body.