Sperm modulate uterine immune parameters relevant to embryo implantation and reproductive success
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE167485 | GEO | 2021/02/25
REPOSITORIES: GEO
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Project description:To examine the effect of seminal fluid on the whole genome expression profile of endometrial tissue following mating, RNA was extracted from endometrial tissue collected 8 h after CBAF1 females were mated with intact Balb/c males and compared to RNA from endometrial tissue of females mated with seminal fluid deficient SVX/VAS Balb/c males. This comparison controlled for ovarian hormone status, exposure to the male and mating activity, and the neuroendocrine response to cervical and vaginal stimulus at mating, so that changes in endometrial gene expression could be attributed specifically to contact with seminal fluid. The endometrial RNA from n=16 individual females was pooled into four independent biological replicates per treatment group (n=4 endometrial samples per replicate) and expression profiles were analyzed by Affymetrix microarray. Seminal fluid exposure induced a clear difference in the profile of genes expressed in the endometrium with a total of 335 genes were differentially regulated with a fold-change greater than 1.5 and p<0.05. Of these, 190 genes were upregulated and 145 genes were downregulated following contact with seminal fluid. Bioinformatics analysis revealed TLR4 signaling as a strongly predicted upstream regulator activated by the differentially expressed genes.Additional experiments confirmed the role of TLR4 with the absence of TLR4 in TLR4 null mice resulting in a failure for seminal fluid to induce endometrial Csf3, Cxcl2, Il6 and Tnf expression. This study provides evidence that TLR4 contributes to seminal fluid modulation of the periconception immune environment. Activation of TLR4 signaling by microbial or endogenous components of seminal fluid is thus implicated as a key element of the female tract response to seminal fluid at the outset of pregnancy in mice.
2015-07-01 | GSE70401 | GEO
Project description:To examine the effect of seminal fluid on the whole genome expression profile of endometrial tissue following mating, RNA was extracted from endometrial tissue collected 8 h after CBAF1 females were mated with intact Balb/c males and compared to RNA from endometrial tissue of females mated with seminal fluid deficient SVX/VAS Balb/c males. This comparison controlled for ovarian hormone status, exposure to the male and mating activity, and the neuroendocrine response to cervical and vaginal stimulus at mating, so that changes in endometrial gene expression could be attributed specifically to contact with seminal fluid. The endometrial RNA from n=16 individual females was pooled into four independent biological replicates per treatment group (n=4 endometrial samples per replicate) and expression profiles were analyzed by Affymetrix microarray. Seminal fluid exposure induced a clear difference in the profile of genes expressed in the endometrium with a total of 335 genes were differentially regulated with a fold-change greater than 1.5 and p<0.05. Of these, 190 genes were upregulated and 145 genes were downregulated following contact with seminal fluid. Bioinformatics analysis revealed TLR4 signaling as a strongly predicted upstream regulator activated by the differentially expressed genes.Additional experiments confirmed the role of TLR4 with the absence of TLR4 in TLR4 null mice resulting in a failure for seminal fluid to induce endometrial Csf3, Cxcl2, Il6 and Tnf expression. This study provides evidence that TLR4 contributes to seminal fluid modulation of the periconception immune environment. Activation of TLR4 signaling by microbial or endogenous components of seminal fluid is thus implicated as a key element of the female tract response to seminal fluid at the outset of pregnancy in mice. Endometrial RNA collected 8 h following coitus from n=16 individual females was pooled into four independent biological replicates per treatment group (n=4 endometrial samples per replicate) and expression profiles were analyzed by Affymetrix microarray. Microarray analysis was performed using Affymetrix Mouse Gene 2.0 ST Arrays at the Adelaide Microarray Centre (Adelaide, Australia). Total RNA (300 ng) was labelled using the GeneChip® WT PLUS Reagent Kit according to the manufacturer’s instructions (Affymetrix, Santa Clara, CA). Microarray data was analyzed using Partek Genomics Suite version 6.6). Briefly, .cel files were imported using RMA background correction, following GC content correction and mean probe summarization. Differentially expressed probes were defined as ≥1.50-fold change, t-test p <0.05. To investigate gene pathways and upstream regulators activated by seminal fluid following mating, differentially expressed genes were analyzed using Ingenuity Pathway Analysis (IPA) version 18488943 Build 308606M (IPA, Ingenuity Systems, Redwood City, CA).
2015-07-01 | E-GEOD-70401 | biostudies-arrayexpress
Project description:Ejaculates contain a diverse mixture of sperm and seminal fluid proteins, the combination of which is crucial to male reproductive success under competitive conditions. Males should therefore tailor the production of different ejaculate components according to their social environment, with particular sensitivity to cues of sperm competition risk (i.e. how likely it is that females will mate promiscuously). Here we test this hypothesis using an established vertebrate model system, the house mouse (Mus musculus domesticus), combining experimental data with a quantitative proteomics analysis of seminal fluid composition. Our study tests for the first time how both sperm and seminal fluid components of the ejaculate are tailored to the social environment.
2015-10-01 | PXD002900 | Pride
Project description:The seminal vesicles are male accessory sex glands that contribute the bulk of the seminal plasma in which mammalian spermatozoa are bathed during ejaculation. In addition to helping convey sperm through the ejaculatory duct, seminal vesicle secretions support sperm survival after ejaculation and influence the female reproductive tract to promote receptivity to pregnancy. Despite its biological importance, there is limited understanding of seminal vesicle fluid (SVF) composition with only 75 proteins having been identified in publicly available proteomic analyses. To address this limitation, here we report new preparative methodology involving sequential solubilization of mouse SVF in guanidine hydrochloride, prior to acetone precipitation and analysis by label-free quantitative mass spectrometry. This strategy identified 126 proteins, including 83 previously undetected in SVF. Members of the seminal vesicle secretory protein family were the most abundant, accounting for 79% of all peptide spectrum matches. Functional analysis of SVF proteins identified inflammation and formation of the vaginal plug as the two most prominent biological processes. Notable additional processes included modulation of sperm function and regulation of the female reproductive tract immune environment. Together, these findings provide a robust methodological framework for future SVF studies and identify novel proteins with the potential to influence both male and female reproductive health.
2022-01-20 | PXD028564 | Pride
Project description:Seminal fluid contains some of the fastest evolving proteins currently known. These seminal fluid proteins (Sfps) play crucial roles in reproduction, such as supporting sperm function, and – particularly in insects – modifying female physiology and behaviour. Identification of Sfps in small animals is challenging, and often relies on samples taken from the female reproductive tract after mating. A key pitfall of this method is that it might miss Sfps that are of low abundance due to dilution in the female-derived sample or rapid processing in females. Here we present a new and complimentary method, which provides added sensitivity to Sfp identification. We applied label-free quantitative proteomics to Drosophila melanogaster male reproductive tissue – where Sfps are unprocessed, and highly abundant – and quantified Sfps before and immediately after mating, to infer those transferred during copulation. We also analysed female reproductive tracts immediately before and after copulation to confirm the presence and abundance of known and candidate Sfps, where possible. Results were cross-referenced with transcriptomic and sequence databases to improve confidence in Sfp detection. Our data was consistent with 124 previously reported Sfps. We found 8 high-confidence novel candidate Sfps, which were both depleted in mated versus unmated males and identified within the reproductive tract of mated but not virgin females. We also identified 31 more candidates that are likely Sfps based on their abundance, known expression and predicted characteristics, and revealed that four proteins previously identified as Sfps are at best minor contributors to the ejaculate. The estimated copy numbers for our candidate Sfps were lower than for previously identified Sfps, supporting the idea that our technique provides a deeper analysis of the Sfp proteome than previous studies. Our results demonstrate a novel, high-sensitivity approach to the analysis of seminal fluid proteomes, whose application will further our understanding of reproductive biology.
2019-08-26 | PXD009451 | Pride
Project description:Sperm competition theory predicts that males should tailor ejaculates according to their social status. Here we test this in a model vertebrate, the house mouse (Mus musculus domesticus), combining experimental data with a quantitative proteomics analysis of seminal fluid composition. Our analyses reveal that both sperm production and the relative production of proteins found in seminal fluid differ according to social dominance. Notably, whereas dominant males produce and ejaculate more sperm, subordinate males produce greater relative amounts of key proteins used in the formation of copulatory plugs. These findings have important implications for understanding the dynamics and outcome of sperm competition.
2021-09-09 | PXD019727 | Pride
Project description:Transcriptional profiling of adult mouse liver tissue comparing offspring derived from sperm and seminal plasma of normal protein diet fed males (controls, NN), sperm and seminal plasma from males fed a low protein diet fed males (LL), sperm from normal protein fed males and seminal plasma from low protein fed males (NL) or sperm from low protein diet fed males and seminal plasma from normal protein diet males (NL). The first letter denotes the diet of the sperm donor and the second letter the diet of the seminal plasma donor. Three-condition experiment: NN vs. LL, NN vs. NL, NN vs. LN. Adult offspring liver tissue. Biological replicates: 7 control (NN), 9 LL, 7 NL and 7 LN. One replicate per array chip.
2022-09-20 | GSE211520 | GEO
Project description:Globally invasive Aedes aegypti mosquitoes disseminate numerous arboviruses that impact human health. One promising method to control Ae. aegypti populations is transinfection with the intracellular bacterium Wolbachia pipientis, a symbiont that naturally infects ~40-52% of insects but is normally absent from Ae. aegypti. Transinfection of Ae. aegypti with the wMel Wolbachia strain induces cytoplasmic incompatibility (CI), allowing infected individuals to rapidly invade native populations. Further, wMel Wolbachia-infected females display refractoriness to medically relevant arboviruses. Thus, wMel Wolbachia-infected Ae. aegypti are being released in several areas to replace native populations, thereby suppressing disease transmission by this species. Wolbachia is reported to have minimal effects on Ae. aegypti fertility, but its influence on other reproductive processes is unknown. Female insects undergo several post-mating physiological and behavioral changes required for optimal fertility. Post-mating responses (PMRs) in female insects are typically elicited by receipt of male seminal fluid proteins (SFPs) transferred with sperm during mating, but can be modified by other factors, such as adult age, nutritional status, and microbiome composition. To assess how Wolbachia infection influences Ae. aegypti female PMRs, we collected wMel Wolbachia-infected Ae. aegypti from the field in Medellín, Colombia and introduced the bacterium into our laboratory strain. We found that Wolbachia influences female fecundity, fertility, and re-mating incidence. Further, we observed that Wolbachia significantly extends longevity of virgin females. Changes in female PMRs are not due to defects in sperm transfer by infected males, or sperm storage by infected females. Using proteomic methods to examine the seminal proteome of infected males, we found that Wolbachia infection has a moderate effect on SFP composition. However, we identified 125 Wolbachia proteins that are paternally transferred to females by infected males. Surprisingly, the CI factor proteins (Cifs), were not detected in the ejaculates of Wolbachia-infected males. Our findings indicate that Wolbachia infection of Ae. aegypti alters female post-mating responses, potentially influencing control programs that utilize Wolbachia-infected individuals.
2023-10-24 | PXD043965 | Pride
Project description:Interactions between sperm and the female reproductive tract (FRT) are critical to fertility, but knowledge of the molecular mechanisms by which the FRT interacts with sperm and influences sperm fate remains limited. Here, we used whole-cell quantitative proteomics to track the protein composition of sperm across three female reproductive tissues (bursa, seminal receptacle, and spermatheca) and three post-mating timepoints (30 minutes, 2 hours, and 4 days). In combination with an update and expansion of the seminal vesicle sperm proteome and sex-specific isotopic labeling to identify female-contributed sperm proteins, our data provide a comprehensive, quantitative analysis of the molecular life history of Drosophila sperm.
2022-03-14 | PXD027807 | Pride
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.
2019-01-03 | PXD011320 | Pride