Gene expression profile of experimentally Deformed wing virus (DWV)-infected honey bees: Naive control vs. DWV-infected pupae
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Honey bee (Apis mellifera) colony., Total RNA from honey bee brains, Brains of infected and control honey bees, Labeled RNA, Hybridized arrays, White eye pupae without manipulation., White-eye pupae, purified DWV particles
ORGANISM(S): Apis mellifera
SUBMITTER: Elke Genersch
PROVIDER: E-MTAB-5070 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Experiment was designed (i) to analyse the strain composition of Deformed wing virus (DWV) populations in covertly and overtly infected honeybees (Apis mellifera) from Varroa-free and Varroa-infested colonies, and (ii) to determine abundance of the DWV strains following direct injection of the DWV preparations from covertly and overtly infected bees to the bee pupae haemolymph in the absence of Varroa destructor mites. Experiment included isolation of DWV preparations from the following bees: covertly-infected bees from Varroa-free colony, covertly infected bees exposed orally to the Varroa-selected DWV strains, and the overtly infected Varroa-exposed bees. Honeybee pupae were experimentally injected with those DWV preparations and sampled 4 days post injection following development of overt DWV infection. A series of the DWV cDNA fragment covering complete DWV genomic RNA sequences were amplified by RT-PCR using RNA extracted from virus preparations and the injected pupae. The cDNA preparations were sequenced using next generation(Illumina HighSeq 2000) paired-end sequencing to obtain data on the DWV strain composition.
2013-09-01 | E-MTAB-1675 | biostudies-arrayexpress
Project description:Experiment was designed to study the effect of Deformed wing virus (DWV) and the mite Varroa destructor on on siRNA and miRNA composition using high-throughput sequencing of small RNA in developing worker honeybee (Apis mellifera). Newly hatched bee larvae (day 3 of bee development) were transferred from a Varroa-free colony with low DWV levels to a Varroa-infested colony with high levels of DWV in bees and Varroa mites. All transferred larvae were receiving the DWV strains present in this Varroa-infested colony with the food delivered by the nurse bees until their capping (day 8). About half of these larvae were capped with Varroa mite and were subjected to the mite piercing and feeding on their haemolymph during pupal development until sampling at purple eye stage (day 14). Exposure to the mite piercing and feeding resulted in about 1000-fold increase of the DWV levels in the majority of the mite-exposed pupae compared to the control pupae and the pupae not exposed to Varroa mites.
2013-09-01 | E-MTAB-1671 | biostudies-arrayexpress
Project description:Experiment was designed to study the effect of Deformed wing virus (DWV) and the mite Varroa destructor on global gene expression using microarray transcriptional profiling in developing worker honeybee (Apis mellifera). Newly hatched bee larvae (day 3 of bee development) were transferred from a Varroa-free colony with low DWV levels to a Varroa-infested colony with high levels of DWV in bees and Varroa mites. All transferred larvae were receiving the DWV strains present in this Varroa-infested colony with the food delivered by the nurse bees until their capping (day 8). About half of these larvae were capped with Varroa mite and were subjected to the mite piercing and feeding on their haemolymph during pupal development until sampling at purple eye stage (day 14). Exposure to the mite piercing and feeding resulted in about 1000-fold increase of the DWV levels in the majority of the mite-exposed pupae compared to the control pupae and the pupae not exposed to Varroa mites.
2013-06-01 | E-MTAB-1285 | biostudies-arrayexpress
Project description:As a matter of fact, honeybees are vital for the pollination of more than 80 crops of agricultural interest. However, population decline has become an important global issue causing significant concerns among agricultural experts and the broader public. For this, parasites are known to be the major culprits responsible for the losses of millions of honeybee colonies so far. Among these parasites, Varroa destructor has been identified as a major cause for global losses in Western honeybee (Apis mellifera) colonies. Hygienic behavior (HB), on the other hand, is a collective response by adult honeybees to defend against parasites and diseases that is known to involve in resistance towards Varroosis. Even with the efforts made to elucidate the molecular mechanism underlying HB, it is still not understood. In our study, we have studied the proteomic correlates to HB using a honeybee line (selected for Varroa-specific HB for over a decade in Germany). We sampled individual worker bees from this line that showed HB after closer infrared video observations and compared the proteomes of their mushroom bodies and antennae with those of workers that came from the same set of colonies but didn't show the behaviour. Furthermore, we compared the pupal hemolymph for worker bees of the selected HB line and a control line using state-of-the art techniques of proteomics. We identified a total of 8609 proteins (covered >55% of the honeybee proteome) from these three honeybee tissues. This is the most comprehensive proteomic study of the honeybee HB to date, and the first to focus on individual bees expressing Varroa-specific HB. These results have significantly advanced our knowledge on the biology underlining HB to a new level. The uniquely found functional classes and pathways by the proteins identified in each tissue suggest that hygienic bees have shaped distinct proteome settings to underpin the HB. Moreover, during analysis of pupal hemolymph proteome, the HB-line has adapted a unique strategy to boost an individual and social immunity and drove pupal organogenesis via energy metabolism and protein biosynthesis. Moreover, in the mushroom bodies of different HB phenotypic worker bees, the hygienic bees have enhanced their neuronal sensitivity to promote the execution of HB by activation of synaptic vesicles and calcium channel activities. Moreover, in the antennae of two HB phenotypic worker bees, the hygienic bees have demonstrated strengthening of their sensitivity associated with olfactory senses and signal transmissions, which is important to input a strong signal to the mushroom bodies and initiate HB. In conclusion, our novel findings have significantly extended our understandings of the molecular mechanisms that underline the HB to combat Varroa infestation. Furthermore, we identified a wide array of novel markers that are useful for accelerating marker associated selection of HB to aid in the natural resistance to a parasite blamed for a global decline in honeybee health.
2016-08-09 | PXD004467 | Pride
Project description:There were important gaps in our knowledge of Israeli acute paralysis virus (IAPV), when IAPV was tightly linked to bee Colony Collapse Disorder (CCD), the mysterious disease that, starting in 2006-2007, has been wiping out honey bees in the US. To fill in these gaps we studied the molecular basis of transmission, pathogenesis, and genetic diversity of IAPV infection in honey bees. We investigated the impact of IAPV infection on colony losses and host transcriptional response to IAPV infections, and exploited the potential of RNAi-based strategies for treating viral diseases in honey bees. Our study clearly shows that IAPV has become established as a persistent infection and is highly prevalent in the honey bee population. The existence of both horizontal and vertical transmission pathways of the virus likely accounts for the high prevalence of IAPV in bees. While IAPV is probably not the only culprit responsible for CCD, its ability to cause increased mortality in honey bees is firmly demonstrated. The phenotypic differences in pathology among different strains of IAPV may be due to their high level of standing genetic variation. The JAK-STAT pathway, along with other signaling events such as mTOR and MAPK pathways, likely involves honey bees’ antiviral immune responses to the IAPV infection. The identification of IAPV-encoded putative suppressor of RNAi and evidence that silencing the RNAi suppressor led to a significant reduction in IAPV replication in infected bees illustrates the therapeutic potential of targeting viral suppressor protein to reduce virus replication. Our study gives direction for developing strategies to reduce colony losses due to viral diseases. Adult worker bees and brood were collected from colonies that were declining and identified with IAPV infections and its control with 6 replications per group.
2014-04-08 | E-GEOD-46278 | biostudies-arrayexpress
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.
2020-10-30 | PXD019392 | Pride
Project description:The genome of the western honey bee (Apis mellifera) harbours ten different major royal jelly protein genes (mrjp1-10) which originate from a single-copy precursor via gene duplication. The evolutionary fate of duplicated genes is eventually determined over time as to result in loss due to pseudogenization, or in preservation due to neo- or sub-functionalization. Both fates were already observed in the mrjp gene cluster, as only mrjp1 - 9 are expressed, whereas mrjp10 was pseudogenized and represents an incomplete gene copy. In contrast, MRJP1 underwent neofunctionalization and developed an essential function within the food jelly of queen larvae, to guaranty the survival of the whole colony. We here show combining quantitative real time PCR with quantitative mass spectrometry that expression of most mrjps (mrjp1-5 and 7) shows an age dependent pattern in worker hypopharyngeal glands as well as in brains. Expression increases after hatching until the nurse bee period and is followed by a decrease in older workers that forage for different plant products. Mrjp6 expression deviates considerably from the expression profiles of the other mrjps and transcript abundance does not correlate with protein amount. Thus, either mrjp6 does fulfil a total different function or it might be on its way to pseudogenization. Furthermore, a tissue-specific function of the proteins MRJP8 and 9 in the hypopharyngeal glands and the brain can be excluded, suggesting a more general physiological than a nutritive function for both gene products.
2019-07-18 | PXD012618 | Pride
Project description:Larvae-Pupae transition flies (Drosophila) were recovered and transport for 3 days at 12-14ºC to arrest development until the launch site, then exposed to RT (18-20ºC) for some hours including the launch and trip to the International Space Station, then pupae were exposed to microgravity in the ISS for 4 days and a half at 22ºC. Finally pupae were fixed on acetone and frozen until recovery on Earth.<br><br><br><br>Four groups of samples: 1 ISS (+ground control) as described, 2 RPM (microgravity simulator on Earth) as described, 3 RPM without constrains (No MAMBA container and only 5 days exposure without cold transport) and 4 centrifuge 10g without constrains control..
2010-07-01 | E-MEXP-2064 | biostudies-arrayexpress
Project description:Bees from 3 unrelated colonies were injected with 1ul PBS extract containing 10^9 genome equivalent of Deformed wIng virus (DWV) and/or fed 10µl sucrose solution containing 10^5 fresh Nosema ceranae spores. Control bees were injected and fed with an equivalent DWV- and Nosema-free extract respectively). Bees were kept in cages of 21 bees (7 from each colony), and each cage was replicated 5 times per each of the 4 treatments). Bess were kept in an incubator at 30°C/50%RH. At day 12 p.i., bees were flash frozen in liquid nitrogen, and stored at -80°C. Bee abdomen RNA was sent to Christina Grozinger lab (Penn State, USA). RNA was pooled for 3 abdomens per replicate for 5 replicates per treatment. Arrays were hybridized in a dye-swap loop design.
2017-03-08 | E-MTAB-5508 | biostudies-arrayexpress
Project description:It is estimated that animals pollinate 87.5% of flowering plants worldwide and that managed honey bees (Apis mellifera) account for 30-50% of this ecosystem service to agriculture. In addition to their important role as pollinators, honey bees are well-established insect models for studying learning and memory, behaviour, caste differentiation, epigenetic mechanisms, olfactory biology, sex determination and eusociality. Despite their importance to agriculture, knowledge of honey bee biology lags behind many other livestock species. In this study we have used scRNA-Seq to map cell types to different developmental stages of the worker honey bee (prepupa at day 11 and pupa at day 15), and sought to determine their gene signatures and thereby provide potential functional annotations for as yet poorly characterized genes. To identify cell type populations we examined the cell-to-cell network based on the similarity of the single-cells’ transcriptomic profiles. Grouping similar cells together we identified 63 different cell clusters of which 15 clusters were identifiable at both stages. To determine genes associated with specific cell populations or with a particular biological process involved in honey bee development, we used gene co-expression analysis. We combined this analysis with literature mining, the honey bee protein atlas and Gene Ontology analysis to determine cell cluster identity. Of the cell clusters identified, 9 were related to the nervous system, 7 to the fat body, 14 to the cuticle, 5 to muscle, 4 to compound eye, 2 to midgut, 2 to hemocytes and 1 to malpighian tubule/pericardial nephrocyte. To our knowledge, this is the first whole single cell atlas of honey bees at any stage of development and demonstrates the potential for further work to investigate their biology of at the cellular level.
2023-07-18 | E-MTAB-13158 | biostudies-arrayexpress