Molecular Diversification of the Seminal Fluid Proteome in a Recently Diverged Passerine Species Pair

Abstract Seminal fluid proteins (SFPs) mediate an array of postmating reproductive processes that influence fertilization and fertility. As such, it is widely held that SFPs may contribute to postmating, prezygotic reproductive barriers between closely related taxa. We investigated seminal fluid (SF) diversification in a recently diverged passerine species pair (Passer domesticus and Passer hispaniolensis) using a combination of proteomic and comparative evolutionary genomic approaches. First, we characterized and compared the SF proteome of the two species, revealing consistencies with known aspects of SFP biology and function in other taxa, including the presence and diversification of proteins involved in immunity and sperm maturation. Second, using whole-genome resequencing data, we assessed patterns of genomic differentiation between house and Spanish sparrows. These analyses detected divergent selection on immunity-related SF genes and positive selective sweeps in regions containing a number of SF genes that also exhibited protein abundance diversification between species. Finally, we analyzed the molecular evolution of SFPs across 11 passerine species and found a significantly higher rate of positive selection in SFPs compared with the rest of the genome, as well as significant enrichments for functional pathways related to immunity in the set of positively selected SF genes. Our results suggest that selection on immunity pathways is an important determinant of passerine SF composition and evolution. Assessing the role of immunity genes in speciation in other recently diverged taxa should be prioritized given the potential role for immunity-related proteins in reproductive incompatibilities in Passer sparrows.

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Sex peptide of Drosophila melanogaster males is a global regulator of reproductive processes in females

Proceedings of The Royal Society B Biological Sciences ◽

10.1098/rspb.2012.1634 ◽

2012 ◽

Vol 279 (1746) ◽

pp. 4423-4432 ◽

Cited By ~ 51

Author(s):

A. Gioti ◽

S. Wigby ◽

B. Wertheim ◽

E. Schuster ◽

P. Martinez ◽

...

Keyword(s):

Drosophila Melanogaster ◽

Sexual Conflict ◽

Seminal Fluid ◽

Nutrient Sensing ◽

Seminal Fluid Proteins ◽

Sex Peptide ◽

Efficient Induction ◽

Transcriptional Changes ◽

Female Behaviour ◽

Reproductive Processes

Seminal fluid proteins (Sfps) alter female behaviour and physiology and can mediate sexual conflict. In Drosophila melanogaster , a single Sfp, the sex peptide (SP), triggers remarkable post-mating responses in females, including altered fecundity, feeding, immunity and sexual receptivity. These effects can favour the evolutionary interests of males while generating costs in females. We tested the hypothesis that SP is an upstream master-regulator able to induce diverse phenotypes through efficient induction of widespread transcriptional changes in females. We profiled mRNA responses to SP in adult female abdomen (Abd) and head+thorax (HT) tissues using microarrays at 3 and 6 h following mating. SP elicited a rich, subtle signature of temporally and spatially controlled mRNAs. There were significant alterations to genes linked to egg development, early embryogenesis, immunity, nutrient sensing, behaviour and, unexpectedly, phototransduction. There was substantially more variation in the direction of differential expression across time points in the HT versus Abd. The results support the idea that SP is an important regulator of gene expression in females. The expression of many genes in one sex can therefore be under the influence of a regulator expressed in the other. This could influence the extent of sexual conflict both within and between loci.

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Interactions between the microbiome and mating influence the female’s transcriptional profile in Drosophila melanogaster

Scientific Reports ◽

10.1038/s41598-020-75156-9 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Sofie Y. N. Delbare ◽

Yasir H. Ahmed-Braimah ◽

Mariana F. Wolfner ◽

Andrew G. Clark

Keyword(s):

Drosophila Melanogaster ◽

Egg Production ◽

Seminal Fluid ◽

Transcript Abundance ◽

Vital Role ◽

Transcriptional Profile ◽

Seminal Fluid Proteins ◽

Two Factors ◽

Immunity Genes ◽

Neuronal Functions

Abstract Drosophila melanogaster females undergo a variety of post-mating changes that influence their activity, feeding behavior, metabolism, egg production and gene expression. These changes are induced either by mating itself or by sperm or seminal fluid proteins. In addition, studies have shown that axenic females—those lacking a microbiome—have altered fecundity compared to females with a microbiome, and that the microbiome of the female’s mate can influence reproductive success. However, the extent to which post-mating changes in transcript abundance are affected by microbiome state is not well-characterized. Here we investigated fecundity and the post-mating transcript abundance profile of axenic or control females after mating with either axenic or control males. We observed interactions between the female’s microbiome and her mating status: transcripts of genes involved in reproduction and genes with neuronal functions were differentially abundant depending on the females’ microbiome status, but only in mated females. In addition, immunity genes showed varied responses to either the microbiome, mating, or a combination of those two factors. We further observed that the male’s microbiome status influences the fecundity of both control and axenic females, while only influencing the transcriptional profile of axenic females. Our results indicate that the microbiome plays a vital role in the post-mating switch of the female’s transcriptome.

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Divergence in sex peptide-mediated female post-mating responses in Drosophila melanogaster

Proceedings of The Royal Society B Biological Sciences ◽

10.1098/rspb.2018.1563 ◽

2018 ◽

Vol 285 (1886) ◽

pp. 20181563 ◽

Cited By ~ 5

Author(s):

Kristina U. Wensing ◽

Claudia Fricke

Keyword(s):

Drosophila Melanogaster ◽

Seminal Fluid ◽

Lifetime Reproductive Success ◽

Fitness Costs ◽

Seminal Fluid Proteins ◽

Sex Peptide ◽

Fitness Measures ◽

Different Populations ◽

Reproductive Processes ◽

Sexually Antagonistic Coevolution

Transfer and receipt of seminal fluid proteins crucially affect reproductive processes in animals. Evolution in these male ejaculatory proteins is explained with post-mating sexual selection, but we lack a good understanding of the evolution of female post-mating responses (PMRs) to these proteins. Some of these proteins are expected to mediate sexually antagonistic coevolution generating the expectation that females evolve resistance. One candidate in Drosophila melanogaster is the sex peptide (SP) which confers cost of mating in females. In this paper, we compared female SP-induced PMRs across three D. melanogaster wild-type populations after mating with SP-lacking versus control males including fitness measures. Surprisingly, we did not find any evidence for SP-mediated fitness costs in any of the populations. However, female lifetime reproductive success and lifespan were differently affected by SP receipt indicating that female PMRs diverged among populations. Injection of synthetic SP into virgin females further supported these findings and suggests that females from different populations require different amounts of SP to effectively initiate PMRs. Molecular analyses of the SP receptor suggest that genetic differences might explain the observed phenotypical divergence. We discuss the evolutionary processes that might have caused this divergence in female PMRs.

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Sperm wars and the evolution of male fertility

Reproduction ◽

10.1530/rep-12-0285 ◽

2012 ◽

Vol 144 (5) ◽

pp. 519-534 ◽

Cited By ~ 210

Author(s):

Leigh W Simmons ◽

John L Fitzpatrick

Keyword(s):

Sperm Competition ◽

Swimming Speed ◽

Male Fertility ◽

Seminal Fluid ◽

Future Research ◽

Sperm Length ◽

Seminal Fluid Proteins ◽

Control Male ◽

Form And Function ◽

And Function

Females frequently mate with several males, whose sperm then compete to fertilize available ova. Sperm competition represents a potent selective force that is expected to shape male expenditure on the ejaculate. Here, we review empirical data that illustrate the evolutionary consequences of sperm competition. Sperm competition favors the evolution of increased testes size and sperm production. In some species, males appear capable of adjusting the number of sperm ejaculated, depending on the perceived levels of sperm competition. Selection is also expected to act on sperm form and function, although the evidence for this remains equivocal. Comparative studies suggest that sperm length and swimming speed may increase in response to selection from sperm competition. However, the mechanisms driving this pattern remain unclear. Evidence that sperm length influences sperm swimming speed is mixed and fertilization trials performed across a broad range of species demonstrate inconsistent relationships between sperm form and function. This ambiguity may in part reflect the important role that seminal fluid proteins (sfps) play in affecting sperm function. There is good evidence that sfps are subject to selection from sperm competition, and recent work is pointing to an ability of males to adjust their seminal fluid chemistry in response to sperm competition from rival males. We argue that future research must consider sperm and seminal fluid components of the ejaculate as a functional unity. Research at the genomic level will identify the genes that ultimately control male fertility.

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BMP signaling inhibition in Drosophila secondary cells remodels the seminal proteome and self and rival ejaculate functions

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1914491116 ◽

2019 ◽

Vol 116 (49) ◽

pp. 24719-24728 ◽

Cited By ~ 2

Author(s):

Ben R. Hopkins ◽

Irem Sepil ◽

Sarah Bonham ◽

Thomas Miller ◽

Philip D. Charles ◽

...

Keyword(s):

Seminal Fluid ◽

Cell Types ◽

Bmp Signaling ◽

Secretory Cells ◽

Normal Female ◽

Seminal Fluid Proteins ◽

Accessory Glands ◽

Offspring Production ◽

Morphogenetic Protein ◽

And Function

Seminal fluid proteins (SFPs) exert potent effects on male and female fitness. Rapidly evolving and molecularly diverse, they derive from multiple male secretory cells and tissues. In Drosophila melanogaster, most SFPs are produced in the accessory glands, which are composed of ∼1,000 fertility-enhancing “main cells” and ∼40 more functionally cryptic “secondary cells.” Inhibition of bone morphogenetic protein (BMP) signaling in secondary cells suppresses secretion, leading to a unique uncoupling of normal female postmating responses to the ejaculate: refractoriness stimulation is impaired, but offspring production is not. Secondary-cell secretions might therefore make highly specific contributions to the seminal proteome and ejaculate function; alternatively, they might regulate more global—but hitherto undiscovered—SFP functions and proteome composition. Here, we present data that support the latter model. We show that in addition to previously reported phenotypes, secondary-cell-specific BMP signaling inhibition compromises sperm storage and increases female sperm use efficiency. It also impacts second male sperm, tending to slow entry into storage and delay ejection. First male paternity is enhanced, which suggests a constraint on ejaculate evolution whereby high female refractoriness and sperm competitiveness are mutually exclusive. Using quantitative proteomics, we reveal changes to the seminal proteome that surprisingly encompass alterations to main-cell–derived proteins, indicating important cross-talk between classes of SFP-secreting cells. Our results demonstrate that ejaculate composition and function emerge from the integrated action of multiple secretory cell types, suggesting that modification to the cellular make-up of seminal-fluid-producing tissues is an important factor in ejaculate evolution.

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BMP-signalling inhibition in Drosophila secondary cells remodels the seminal proteome, and self and rival ejaculate functions

10.1101/741587 ◽

2019 ◽

Author(s):

Ben R. Hopkins ◽

Irem Sepil ◽

Sarah Bonham ◽

Thomas Miller ◽

Philip D. Charles ◽

...

Keyword(s):

Seminal Fluid ◽

Cell Types ◽

Secretory Cells ◽

Normal Female ◽

Seminal Fluid Proteins ◽

Accessory Glands ◽

Offspring Production ◽

Use Efficiency ◽

Bmp Signalling ◽

And Function

ABSTRACTSeminal fluid proteins (SFPs) exert potent effects on male and female fitness. Rapidly evolving and molecularly diverse, they derive from multiple male secretory cells and tissues. In Drosophila melanogaster, most SFPs are produced in the accessory glands, which are composed of ∼1000 fertility-enhancing ‘main cells’ and ∼40, more functionally cryptic, ‘secondary cells’. Inhibition of BMP-signalling in secondary cells suppresses secretion, leading to a unique uncoupling of normal female post-mating responses to the ejaculate: refractoriness stimulation is impaired, but offspring production is not. Secondary cell secretions might therefore make a highly specific contribution to the seminal proteome and ejaculate function; alternatively, they might regulate more global – but hitherto-undiscovered – SFP functions and proteome composition. Here, we present data that supports the latter model. We show that in addition to previously reported phenotypes, secondary cell-specific BMP-signalling inhibition compromises sperm storage and increases female sperm use efficiency. It also impacts second male sperm, tending to slow entry into storage and delay ejection. First male paternity is enhanced, which suggests a novel constraint on ejaculate evolution whereby high female refractoriness and sperm competitiveness are mutually exclusive. Using quantitative proteomics, we reveal a mix of specific and widespread changes to the seminal proteome that surprisingly encompass alterations to main cell-derived proteins, indicating important cross-talk between classes of SFP-secreting cells. Our results demonstrate that ejaculate composition and function emerge from the integrated action of multiple secretory cell-types suggesting that modification to the cellular make-up of seminal fluid-producing tissues is an important factor in ejaculate evolution.

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Interactions between the microbiome and mating influence the female’s transcriptional profile in Drosophila melanogaster

10.1101/2020.05.30.125427 ◽

2020 ◽

Cited By ~ 1

Author(s):

Sofie Y. N. Delbare ◽

Yasir H. Ahmed-Braimah ◽

Mariana F. Wolfner ◽

Andrew G. Clark

Keyword(s):

Drosophila Melanogaster ◽

Egg Production ◽

Seminal Fluid ◽

Transcript Abundance ◽

Vital Role ◽

Transcriptional Profile ◽

Seminal Fluid Proteins ◽

Two Factors ◽

Immunity Genes ◽

Neuronal Functions

AbstractDrosophila melanogaster females undergo a variety of post-mating changes that influence their activity, feeding behavior, metabolism, egg production and gene expression. These changes are induced either by mating itself or by sperm or seminal fluid proteins. In addition, studies have shown that axenic females—those lacking a microbiome—have altered fecundity compared to females with a microbiome, and that the microbiome of the female’s mate can influence reproductive success. However, the extent to which post-mating changes in transcript abundance are affected by microbiome state is not well-characterized. Here we investigated fecundity and the post-mating transcript abundance profile of axenic or control females after mating with either axenic or control males. We observed interactions between the female’s microbiome and her mating status: transcripts of genes involved in reproduction and genes with neuronal functions were differentially abundant depending on the females’ microbiome status, but only in mated females. In addition, immunity genes showed varied responses to either the microbiome, mating, or a combination of those two factors. We further observed that the male’s microbiome status influences the fecundity of both control and axenic females, while only influencing the transcriptional profile of axenic females. Our results indicate that the microbiome plays a vital role in the post-mating switch of the female’s transcriptome.

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Reproductive functions and genetic architecture of the seminal fluid and sperm proteomes of the mosquito Aedes aegypti

10.1101/405431 ◽

2018 ◽

Cited By ~ 2

Author(s):

Ethan C. Degner ◽

Yasir H. Ahmed-Braimah ◽

Kiril Borziak ◽

Mariana F. Wolfner ◽

Laura C. Harrington ◽

...

Keyword(s):

Aedes Aegypti ◽

Seminal Fluid ◽

Protein Translation ◽

Chromosome 1 ◽

Composition Analysis ◽

Seminal Fluid Proteins ◽

Accessory Glands ◽

Significant Enrichment ◽

Intimate Knowledge ◽

Reproductive Processes

AbstractThe yellow fever mosquito, Aedes aegypti, transmits several viruses, including dengue, Zika, and chikungunya. Some proposed efforts to control this vector involve manipulating reproduction to suppress wild populations or replacing them with disease-resistant mosquitoes. The design of such strategies requires an intimate knowledge of reproductive processes, yet our basic understanding of reproductive genetics in this vector remains largely incomplete. To accelerate future investigations, we have comprehensively catalogued sperm and seminal fluid proteins (SFPs) transferred to females in the ejaculate using tandem mass spectrometry. By excluding female-derived proteins using an isotopic labelling approach, we identified 870 sperm proteins and 280 seminal fluid proteins. Functional composition analysis revealed parallels with known aspects of sperm biology and SFP function in other insects. To corroborate our proteome characterization, we also generated transcriptomes for testes and the male accessory glands—the primary contributors to Ae. aegypti sperm and seminal fluid, respectively. Differential gene expression of accessory glands from virgin and mated males suggests that protein translation is upregulated post-mating. Several SFP transcripts were also modulated after mating, but >90% remained unchanged. Finally, a significant enrichment of SFPs was observed on chromosome 1, which harbors the male sex determining locus in this species. Our study provides a comprehensive proteomic and transcriptomic characterization of ejaculate production and composition and thus provides a foundation for future investigations of Ae. aegypti reproductive biology, from functional analysis of individual proteins to broader examination of reproductive processes.

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