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Proteomics on spermathecal fluid of BC queens

ABSTRACT: We sampled honey bee queens from beekeepers throughout British Columbia which were rated as either 'failed' or 'healthy' by beekeepers. We performed shot-gun proteomics on the spermathecal fluid (sperm removed). Proteins were reduced, alkylated, and digested with Lys-C for 4 h in urea digestion buffer. The sample was then diluted 4x with ammonium bicarbonated and digested overnight with trypsin. 2 ug of peptides were analyzed by an EASY-nLC 1000 liquid chromatography system coupled to a Bruker Impact II mass spectrometer. Data were searched with MaxQuant and the zipped txt folder output is provided.

INSTRUMENT(S): Bruker Impact II Q-TOF

ORGANISM(S): Apis Mellifera

SUBMITTER: Leonard Foster

PROVIDER: MSV000085056 | MassIVE | Wed Mar 04 19:39:00 GMT 2020

SECONDARY ACCESSION(S): PXD017886

REPOSITORIES: MassIVE

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Candidate stress biomarkers for queen failure diagnostics.

McAfee Alison A Milone Joseph J Chapman Abigail A Foster Leonard J LJ Pettis Jeffery S JS Tarpy David R DR

BMC genomics 20200820 1

<h4>Background</h4>Queen failure is a persistent problem in beekeeping operations, but in the absence of overt symptoms it is often difficult, if not impossible, to ascertain the root cause. Stressors like heat-shock, cold-shock, and sublethal pesticide exposure can reduce stored sperm viability and lead to cryptic queen failure. Previously, we suggested candidate protein markers indicating heat-shock in queens. Here, we further investigate these heat-shock markers and test new stressors to iden ...[more]

PMID: 32819278

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Proteomics on the ovaries of failed and healthy honey bee queens

Project description:We sampled honey bee queens from beekeepers throughout British Columbia which were rated as either 'failed' or 'healthy' by beekeepers. We performed shot-gun proteomics on the ovaries. Proteins were reduced, alkylated, and digested with Lys-C for 4 h in urea digestion buffer. The sample was then diluted 4x with ammonium bicarbonated and digested overnight with trypsin. Peptides were analyzed by an EASY-nLC 1000 liquid chromatography system coupled to a Bruker Impact II mass spectrometer. Data were searched with MaxQuant and the zipped txt folder output is provided.

2021-04-02 | MSV000087150 | MassIVE

Atta columbica seminal fluid and spermathecal fluid proteome

Project description:Mature males and virgin queens from Atta colombica ants were collected in panama, and their seminal fluid (SF) and spermathecal fluid (SpF), respectively, were extracted and analysed by shot-gun proteomics. For this, using a pooled SF sample of 25 males from five colonies, squeezing their abdomens to collect 2-4ul of ejaculate per male from the extended endophallus. The SF supernatant was collected from the pooled ejaculate after 2 centrifugations for10 min at 13,000xg. The Spf was obtained form 4 different colonies collecting 10 virgin queens per colony and “artificially inseminating them with 10ul of Hayes. All 40 queens were allowed to recover for 12 hours before retrieving the content by dissecting and rupturing the spermathecal in Hayes. The samples were vortexed for 3 min, followed by centrifugation for 3 min at 13,500 xg to separate SpF from tissue.

2019-01-03 | PXD011306 | Pride

Seminal fluid compromises visual perception in honeybee queens reducing their survival during additional mating flights

Project description:Queens of social insects make all mate-choice decisions on a single day, except in honeybees whose queens can conduct mating flights for several days even when already inseminated by a number of drones. Honeybees therefore appear to have a unique, evolutionarily derived form of sexual conflict: a queen’s decision to pursue risky additional mating flights is driven by later-life fitness gains from genetically more diverse worker-offspring but reduces paternity shares of the drones she already mated with. We used artificial insemination, RNA-sequencing and electroretinography to show that seminal fluid induces a decline in queen vision by perturbing the phototransduction pathway within 24-48 hours. Follow up field trials revealed that queens receiving seminal fluid flew two days earlier than sister queens inseminated with saline, and failed more often to return. These findings are consistent with seminal fluid components manipulating queen eyesight to reduce queen promiscuity across mating flights.

2019-09-10 | GSE127185 | GEO

Proteome of spermathecal fluid and hemolymph in Lasius japonicus queens

Project description:To reveal the molecular basis of the long-term sperm storage mechanisms in ant queens, protein profiles enriched in the spermathecal fluid relative to the hemolymph were identified in Lasius japonicus using data-independent acquisition (DIA)-based quantitative proteomics technology.

2024-02-01 | PXD041534 | JPOST Repository

Seminal fluid components alone stimulate post-mating changes in honey bee queens (Apis mellifera)

Project description:Honey bee queens undergo dramatic behavioral (e.g., reduced sexual receptivity), physiological (e.g., ovary activation, ovulation, and modulation of pheromone production) and molecular changes after they complete mating. To elucidate how queen post-mating changes are influenced by seminal fluid, a non-spermatozoa-containing component of semen, we injected queens with semen or seminal fluid alone. We assessed queen sexual receptivity, ovary development, worker retinue response (which is influenced by queen pheromone production), and transcriptional changes in queen abdominal fat body and brain tissues. Injection with either seminal fluid or semen resulted in decreased sexual receptivity, increased attractiveness of queens to workers, and altered expression of several genes that are also regulated in naturally mated queens. The post-mating and transcriptional changes of queens receiving seminal fluid were not significantly different from queens treated with seminal fluid, suggesting that components in seminal fluid, such as seminal fluid proteins, are largely responsible for stimulating post-mating changes in queens.

2020-07-27 | GSE145395 | GEO

Comparison of fire ant adult queens with different supergene genotypes (SB/SB and SB/Sb) and different age classes (1-day-old virgins, 11-day-old virgins and reproductive queens) from 2008 and 2009

Project description:In the fire ant Solenopsis invicta, a colony queen number is determined by the founding queen's genotypes at the 13 Mb supergene with the non-recombining variants SB and Sb. Single-queen colonies are always headed by SB/SB queens while multiple-queens colonies are always headed by SB/Sb queens. The two variants of the supergene, SB and Sb are completely linked to the two alleles (B and b) of the gene Gp-9. SB/SB and SB/Sb queens differ in many physiological traits including their maturation rate and odor. To explain why SB/SB and SB/Sb queens have different odors, and why SB/SB virgins mature faster and accumulate more fat, we measured expression of ~6000 genes in virgin queens 1 and 11 days after eclosion and in reproductive queens. Keywords: fire ants, Solenopsis invicta, Supergene, queen, Gp-9, social form, maturation, fat storage, queen odor, cuticular hydrocarbon, worker discrimination, monogyne, polygyne, transposon, chemical signaling

2012-11-07 | GSE42062 | GEO

Effects of mating on gene expression profiles of mature ant queens

Project description:Will be added/updated once the manuscript is finalized. Cardiocondyla obscurior queens. Three treatments: virgin queens, queens mated by real males, queens sham-mated (by sterile males). Queens collected 1 week and 8 week after mating. seven loops for queens collected 1 week after mating; nine loops for queens collected 8 weeks after mating. Five direct comparisons (with dye-swaps... so 10 arrays) were done of between 1 week and 8 week samples of queens mated by real males. Each sample is RNA from two queens (from different colonies). Samples were hybridized against Solenopsis invicta microarrays (signal was detectable for most clones!)

2013-10-26 | E-GEOD-24953 | biostudies-arrayexpress

Costs of reproduction and their effect on age-related gene expression in Bombus terrestris queens

Project description:Our two main aims were 1) to isolate age-related changes in gene expression in queens of the eusocial bumble bee, Bombus terrestris; and 2) to determine whether experimentally increasing the costs of reproduction (by removing eggs) caused changes in age-related gene expression in these queens. To address these aims we extracted RNA from three key tissues (brain, fat body and ovary) from queens at two time points (10% and 60% mortality phases). Each of these queens had experienced one of two treatments: an egg removal (R) treatment (to increase the costs of reproduction) and an egg removal and replacement (C) treatment (to control for the effects of disturbance caused by egg removal).

2023-06-06 | GSE172422 | GEO

Timecourse of gene expression after oprhaning of virgin Solenopsis invicta fire ant queens

Project description:In species with social hierarchies, the death of dominant individuals typically upheaves the social hierarchy and provides an opportunity for subordinate individuals to become reproductives. Such a phenomenon occurs in the monogyne form of the fire ant, Solenopsis invicta, where colonies typically contain a single wingless reproductive queen, thousands of workers and hundreds of winged non-reproductive virgin queens. Upon the death of the mother queen, many virgin queens shed their wings and initiate reproductive development instead of departing on a mating flight. Workers progressively execute almost all of them over the following weeks. To identify the molecular changes that occur in virgin queens as they perceive the loss of their mother queen and begin to compete for reproductive dominance, we collected virgin queens before the loss of their mother queen, six hours after orphaning and 24 hours after orphaning. Their RNA was extracted and hybridized against microarrays to examine the expression levels of approximately 10,000 genes. We identified 297 genes that were consistently differentially expressed after orphaning. These include genes that are putatively involved in the signaling and onset of reproductive development, as well as genes underlying major physiological changes in the young queens.

2010-02-01 | GSE19721 | GEO

The outcome of a competition for reproduction between fire ant queens is linked to their gene expression profiles

Project description:In many animals living in groups the reproductivestatus of individuals is determined by their social status. In specieswith social hierarchies, the death of dominant individuals typicallyupheaves the social hierarchy and provides an opportunity for subordinateindividuals to improve their social status. Such a phenomenon occursin the monogyne form of the fire ant\emph{, Solenopsis invicta, }wherecolonies typically contain a single wingless reproductive queen, thousandsof workers and hundreds of winged non-reproductive virgin queens.Upon the death of the mother queen, many virgin queens shed theirwings and initiate reproductive development instead of departing ona mating flight. Workers progressively execute almost all of themover the following weeks. The workers base their collective decisionon pheromonal cues associated with the onset of reproductive developmentof the virgin queens which occurs after orphaning. To examine the factors that determine which virgin queens are executed and which survive, we set up artificial competitions between queens from different colonies. Using microarrays, we found that queens from winning colonies showed higher mitochondrial as well as organ development activities 24 hours after orphaning than did queens from colonies that lost the competitions. Furthermore, queens from colonies where queens shed their wings faster after orphaning were more likely to survive competitions. Finally, higher wing shedding speed is linked to higher mitochondrial activity.

2013-12-14 | GSE26039 | GEO

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