Proteomic analysis of queen cell wax and cell wax
Ontology highlight
ABSTRACT:
ORGANISM(S): Apis Mellifera
SUBMITTER: 
Yu Fang
PROVIDER: PXD036838 | iProX | Mon Sep 19 00:00:00 BST 2022
REPOSITORIES: iProX
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Project description:Analysis and comparison of protein composition between cell wax and queen cellwax
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Project description:Honeybee health is a major ecological, agricultural and societal concern due to the critical role of these insects for plant reproduction and the massive losses of colonies observed within the last decade. The search for abiotic (e.g. pesticides) and biotic (e.g. pathogens) stressors is essential for understanding bee declines and design protection plans. Flagellated Trypanosomatid protozoan parasites and particularly Lotmaria passim, are widely distributed in honeybee colonies and have been associated with colony losses. However, little is known about their life cycles, routes for transmission and the strategies for survival inside and outside their hosts. Here we describe the proteome of L. passim Extracellular polymeric substances linked to secretion of Extracellular vesicles using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The EPS-EVs composition will provide clues on how these pathogens survive and spread in apiaries worldwide.
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Project description:In the stem-cell niche, the extracellular matrix (ECM) serves as a structural support that additionally provides stem cells with signals that contribute to the regulation of stem-cell function, via reciprocal interactions between cells and components of the ECM. Recently, cell-derived ECMs have emerged as in vitro cell culture substrates to better recapitulate the native stem-cell microenvironment outside the body. Significant changes in cell number, morphology and function have been observed when mesenchymal stem cells (MSC) were cultured on ECM substrates as compared to standard tissue-culture polystyrene (TCPS). As select ECM components are known to regulate specific stem-cell functions, a robust characterization of cell-derived ECM proteomic composition is critical to better comprehend the role of the ECM in directing cellular processes. Here, we characterized and compared the protein composition of ECM produced in vitro by bone marrow-derived MSC, adipose-derived MSC and neonatal fibroblasts isolated from different donors, employing quantitative proteomic methods. Each cell-derived ECM displayed a specific and unique matrisome signature, yet they all shared a common set of proteins. We evaluated the biological response of cells cultured on the different matrices and compared them to cells on standard TCPS. The matrices lead to differential survival and gene-expression profiles among the cell types and as compared to TCPS, indicating that the cell-derived ECMs influence each cell type in a different manner. This general approach to understanding the protein composition of different tissue-specific and cell-derived ECM will inform the rational design of defined systems and biomaterials that recapitulate critical ECM signals for stem-cell culture and tissue engineering.
2017-03-07 | GSE94667 | GEO
Project description:The honeybee (Apis mellifera) is a well-known eusocial insect. In honeybee colonies, thousands of sterile workers, including nurse and forager bees, perform various tasks within or outside the hive, respectively. The queen is the only fertile female and is responsible for reproduction. The queen and workers share similar genomes, but occupy different caste statuses. We established single-cell transcriptomic atlases of brains from queens and worker subcastes, and identified five major cell groups: Kenyon, optic lobe, olfactory projection, glial, and hemocyte cells. By dividing Kenyon and glial cells into multiple subtypes based on credible markers, we observed that vitellogenin (vg) was highly expressed in specific glial-cell subtypes in brains of queens. Knockdown of vg at the early larval stage significantly suppressed the development into adult queens. We demonstrate vg expression as a "molecular signature" for the queen caste, and suggest involvement of vg in regulating caste differentiation.
2022-07-11 | GSE184507 | GEO
Project description:The honeybee brain is comprised of a nervous system that sufficiently regulates this life transition. Knowledge about how protein phosphorylation functions in regards to the neurobiological activities in the honeybee brain to drive the age-specific labor division is still lacking. Protein phosphorylation, the most common post-translational modification (PTM), is a key switch for rapid on-off control of signaling cascades that regulate cell differentiation and development, enzyme activity and metabolic maintenance in living cells. A fundamental mechanism for regulating signaling network and protein activity is the covalent PTM of serine (Ser), threonine (Thr), and tyrosine (Tyr) residues with phosphate. Fortunately, because of advances in phosphopeptide enrichment and improvements in mass spectrometry (MS) instrumentation and methods, phosphoproteomics has enabled large-scale identification of protein phosphorylation sites and phosphorylation networks in biological samples. Although the proteome has been mapped in the brain of nurse and forager bees, knowledge about age-specific effects of phosphorylation regulation on proteins in the honeybee brain is still lacking. Moreover, information in regards to the honeybee phosphoproteome is also very limited. Only very recently, in-depth phosphoproteomics analyses of protein phosphorylation networks in the hypopharyngealgland of the honeybee have been reported. Although the phosphoproteome analyses during the development of brood and salivary glands has been reported, only very limited proteins were phosphorylated and phosphorylation sites of those phosphoproteins were not assigned. Therefore, a comprehensive characterization of phosphoproteomics and changes in the brains of nurse and forager bees is key to understand the phosphorylation events underlying age-specific physiology to achieve the completion of biological missions in this well-organized social community of the honeybee. Honeybee (A. m. ligustica) colonies used for sampling were raised at the apiary of the Institute of Apicultural Research, Chinese Academy of Agricultural Science, Beijing. Newly emerged (<12 h after emergence) worker bees were marked on their thoraxes and placed back into the colonies to develop and then the marked nurse and forager bees were collected on days 10 and 20, respectively. There were 150 bees sampled from each of the five colonies which have queens at the same age. In brief, for each time point, worker bees were sampled from five colonies, and pooled all samples for further analysis. This procedure was repeated three times, so that we finally ended up with three independent biological replicates per time point, each consisting of 150 honeybees. Then their brains were dissected, and the brain samples were pooled and stored at −80 °C for further analysis. All the colonies were managed with almost identical population, food, and brood during the nectar flow of chaste berry (Vitexnegundo L.) in June.
2018-10-26 | PXD003757 | Pride
Project description:Sex determination in the honeybee (Apis mellifera) is governed by the queen-controlled unfertilization or fertilization of embryo, though the mechanisms of determination are poorly understood. Here, we obtained the transcriptomes from individual worker and drone embryo during the embryonic development (day 1 to day 3). We show that transcriptional difference between worker and drone embryo is very small during the first day of hatching, during which sex-determinant gene csd expresses similarly. Differential transcription between worker and drone embryo bursts at day 2, among which csd is induced in worker embryo at day 2 and sex-lethal gene sxl is repressed in male embryo. An unexpected global regulation of alternative splicing accompanies the honeybee embryonic development, and male and worker embryo show distinct regulatory patterns and mechanisms. This study suggests the honeybee sex determination is more globally controlled at both the transcriptional and alternative splicing levels.
2021-07-27 | GSE101741 | GEO
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:E-cadherin upregulation is an early event of reprogramming of fibroblasts to induce pluripotent stem cells (iPS). Knocking down of E-cadherin by shRNA impairs iPS generation, though some colonies with great morphorlogical difference to shRNA control colonies remain. To illustrate the molecular and functional difference between shECAD iPS clones and shRNA control iPS clones, three respective iPS clones (shECAD 4,8,9 and Ctrl 2,3,4) were derived and DNA microarrays were run to analyze the transcriptional profile of these clones.
2010-06-01 | GSE21062 | GEO
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.
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