Dataset Information

Atta columbica seminal fluid and spermathecal fluid proteome

ABSTRACT: 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.

INSTRUMENT(S): 6550 iFunnel Q-TOF LC/MS

ORGANISM(S): Atta Columbica

TISSUE(S): Seminal Vesicle Fluid, Bodily Fluid, Spermatheca

SUBMITTER: Julia Grassl

LAB HEAD: Prof Boris Baer

PROVIDER: PXD011306 | Pride | 2019-01-03

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 98

Publications

Protein-Level Interactions as Mediators of Sexual Conflict in Ants.

Dosselli Ryan R Grassl Julia J den Boer Susanne P A SPA Kratz Madlen M Moran Jessica M JM Boomsma Jacobus J JJ Baer Boris B

Molecular & cellular proteomics : MCP 20181231 Suppl 1

All social insects with obligate reproductive division of labor evolved from strictly monogamous ancestors, but multiple queen-mating (polyandry) arose <i>de novo</i>, in several evolutionarily derived lineages. Polyandrous ant queens are inseminated soon after hatching and store sperm mixtures for a potential reproductive life of decades. However, they cannot re-mate later in life and are thus expected to control the loss of viable sperm because their lifetime reproductive success is ultimately ...[more]

PMID: 30598476

Similar Datasets

Project description:Polyandrous ant queens are inseminated early in their life and store sperm mixtures for a potential reproductive life of decades. However, they cannot re-mate later in life and are thus expected to control the loss of viable sperm because their life-time reproductive success is ultimately limited by the viability of sperm obtained in their single reproductive event. In the leaf-cutting ant Atta colombica, we have previously shown that sperm survival is lowered when getting in contact with seminal fluid of other males, and remains stable when in contact with secretions of the queen sperm storage organ (spermatheca). Here we aim to resolve the main protein-level interactions that mediate sperm competition dynamics and sperm preservation. We used artificial insemination and DIGE-based proteomics to identify proteomic changes when seminal fluid is exposed to spermathecal fluid. Seminal fluid was collected from mature males during field season experiments in Panama, by gently squeezing males’ abdomen until the ejaculate came out. Ejaculates from several males were kept together in ice and then centrifuged for 10 minutes at 13,500 g, the supernatant collected in a separate tube and then centrifuged again. The resulting supernatant was then transferred to another tube and stored at -20° C until further use. Four biological replicates of SF were collected from 4 colonies, each consisting of 400 µl SF from approximately 350 males. From each biological replicate three 100 µl aliquots were retrieved, and assigned to one of the following treatments: (1) un-inseminated controls, (2) inseminated SF retrieved after 30 min, and (3) inseminated SF retrieved after 12 h (Fig 1). Next, each of those aliquots was used to artificially inseminate 10 virgin queens (10 µl of aliquot per queen), for a total of 80 queens inseminated. Queens were allowed to recover for 30 min or 12 h before dissecting them to obtain the spermathecal content. All these spermathecal contents were then pooled per treatment and replicate, resulting in a total of 12 samples (3 treatments – un-inseminated SF, inseminated SF retrieved after 30 min inside the queen, and inseminated SF retrieved after 12 hours inside the queen - with 4 biological replicates). Proteins were precipitated in each sample by adding 4 volumes of ice-cold acetone for 4 hours at -20°C and centrifuged at 14,000 g for 10 minutes, after which supernatants were discarded and the protein pellets were stored at -80°C until further use. All of those 12 samples entered the DIGE analysis.

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. Eight competitions were initially conducted between queens form pairs of colonies. Six competitions clearly identified one winning colony and one losing colony. We thus have microarray data for six competitions (ie biological replicates), with one winning and one losing colony within each competition. Thus 12 colonies total. For each colony, RNA extracted from five queens was pooled, and hybridized against an unrelated common reference RNA that had been made from a pool of many S. invicta individuals of all castes and developmental stages. Lab work was conducted in blocks (by competition) with randomized order within competition to avoid introducing bias. No technical replication was performed.

			Action	DRS
	ACOL SF merged.pride.mgf.gz	Mgf
	ACOLSFmerged.mgf	Mgf
	COL13.mzdata.xml	Xml
	COL14.mzdata.xml	Xml
	COL15.mzdata.xml	Xml