scholarly journals Quantitative proteomics identification of seminal fluid proteins in male Drosophila melanogaster

2018 ◽  
Author(s):  
Irem Sepil ◽  
Ben R Hopkins ◽  
Rebecca Dean ◽  
Marie-Laëtitia Thézénas ◽  
Philip D Charles ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Quantitative Proteomics ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
High Sensitivity ◽  
Female Reproductive Tract ◽  
Label Free ◽  
Seminal Fluid Proteins ◽  
Copy Numbers ◽  
Before And After

AbstractSeminal 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.

scholarly journals Differences in Postmating Transcriptional Responses between Conspecific and Heterospecific Matings in Drosophila

2020 ◽  
Author(s):  
Yasir H Ahmed-Braimah ◽  
Mariana F Wolfner ◽  
Andrew G Clark
Keyword(s):  
Reproductive Tract ◽  
Evolutionary Dynamics ◽  
Seminal Fluid ◽  
Female Reproductive Tract ◽  
Additional Stress ◽  
Reproductive Proteins ◽  
Transcriptional Responses ◽  
Seminal Fluid Proteins ◽  
Reproductive Genes ◽  
Imd Pathway

Abstract In many animal species, females undergo physiological and behavioral changes after mating. Some of these changes are driven by male-derived seminal fluid proteins and are critical for fertilization success. Unfortunately, our understanding of the molecular interplay between female and male reproductive proteins remains inadequate. Here, we analyze the postmating response in a Drosophila species that has evolved strong gametic incompatibility with its sister species; Drosophila novamexicana females produce only ∼1% fertilized eggs in crosses with Drosophila americana males, compared to ∼98% produced in within-species crosses. This incompatibility is likely caused by mismatched male and female reproductive molecules. In this study, we use short-read RNA sequencing to examine the evolutionary dynamics of female reproductive genes and the postmating transcriptome response in crosses within and between species. First, we found that most female reproductive tract genes are slow-evolving compared to the genome average. Second, postmating responses in con- and heterospecific matings are largely congruent, but heterospecific matings induce expression of additional stress-response genes. Some of those are immunity genes that are activated by the Imd pathway. We also identify several genes in the JAK/STAT signaling pathway that are induced in heterospecific, but not conspecific mating. While this immune response was most pronounced in the female reproductive tract, we also detect it in the female head and ovaries. These results show that the female’s postmating transcriptome-level response is determined in part by the genotype of the male, and that divergence in male reproductive genes and/or traits can have immunogenic effects on females.

scholarly journals Differences in post-mating transcriptional responses between conspecific and heterospecific matings in Drosophila

2020 ◽  
Author(s):  
Yasir H. Ahmed-Braimah ◽  
Mariana F. Wolfner ◽  
Andrew G. Clark
Keyword(s):  
Reproductive Tract ◽  
Evolutionary Dynamics ◽  
Seminal Fluid ◽  
Female Reproductive Tract ◽  
Additional Stress ◽  
Reproductive Proteins ◽  
Transcriptional Responses ◽  
Seminal Fluid Proteins ◽  
Reproductive Genes ◽  
Imd Pathway

AbstractIn many animal species, females undergo physiological and behavioral changes after mating. Some of these changes are driven by male-derived seminal fluid proteins, and are critical for fertilization success. Unfortunately, our understanding of the molecular interplay between female and male reproductive proteins remains superficial. Here we analyze the post-mating response in a Drosophila species that has evolved strong gametic incompatibility with its sister species; D. novamexicana females produce only 1% fertilized eggs in crosses with D. americana males, compared to ~98% produced in within-species crosses. This incompatibility is likely caused by mismatched male and female reproductive molecules. In this study we use short-read RNA sequencing to examine the evolutionary dynamics of female reproductive genes and the post-mating transcriptome response in crosses within and between species. First, we found that most female reproductive tract genes are slow-evolving compared to the genome average. Second, post-mating responses in con- and heterospecific matings are largely congruent, but heterospecific mating induces expression of additional stress-response genes. Some of those are immunity genes that are activated by the Imd pathway. We also identify several genes in the JAK/STAT signaling pathway that are induced in heterospecific, but not conspecific mating. While this immune response was most pronounced in the female reproductive tract, we also detect it in the female head and ovaries. Our results show that the female’s post-mating transcriptome-level response is determined in part by the genotype of the male, and that divergence in male reproductive genes and/or traits can have immunogenic effects on females.

scholarly journals Quantitative Proteomics Identification of Seminal Fluid Proteins in Male Drosophila melanogaster

2018 ◽  
Vol 18 (Supplement 1) ◽  
pp. S46-S58 ◽  
Author(s):  
Irem Sepil ◽  
Ben R. Hopkins ◽  
Rebecca Dean ◽  
Marie-Laëtitia Thézénas ◽  
Philip D. Charles ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Quantitative Proteomics ◽  
Seminal Fluid ◽  
Seminal Fluid Proteins

scholarly journals The Drosophila melanogaster Seminal Fluid Protein Acp62F Is a Protease Inhibitor That Is Toxic Upon Ectopic Expression

Genetics ◽  
2002 ◽  
Vol 160 (1) ◽  
pp. 211-224
Author(s):  
Oliver Lung ◽  
Uyen Tram ◽  
Casey M Finnerty ◽  
Marcie A Eipper-Mains ◽  
John M Kalb ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Protease Inhibitor ◽  
Life Span ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
Ectopic Expression ◽  
Circulatory System ◽  
Sperm Storage ◽  
Egg Laying ◽  
Seminal Fluid Proteins

Abstract Drosophila melanogaster seminal fluid proteins stimulate sperm storage and egg laying in the mated female but also cause a reduction in her life span. We report here that of eight Drosophila seminal fluid proteins (Acps) and one non-Acp tested, only Acp62F is toxic when ectopically expressed. Toxicity to preadult male or female Drosophila occurs upon one exposure, whereas multiple exposures are needed for toxicity to adult female flies. Of the Acp62F received by females during mating, ~10% enters the circulatory system while ~90% remains in the reproductive tract. We show that in the reproductive tract, Acp62F localizes to the lumen of the uterus and the female's sperm storage organs. Analysis of Acp62F's sequence, and biochemical assays, reveals that it encodes a trypsin inhibitor with sequence and structural similarities to extracellular serine protease inhibitors from the nematode Ascaris. In light of previous results demonstrating entry of Acp62F into the mated female's hemolymph, we propose that Acp62F is a candidate for a molecule to contribute to the Acp-dependent decrease in female life span. We propose that Acp62F's protease inhibitor activity exerts positive protective functions in the mated female's reproductive tract but that entry of a small amount of this protein into the female's hemolymph could contribute to the cost of mating.

Post-Copulatory Sexual Selection

2021 ◽  
Author(s):  
Patricia L.R. Brennan ◽  
Dara N. Orbach
Keyword(s):  
Sexual Selection ◽  
Sperm Competition ◽  
Female Choice ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
Fertilization Success ◽  
Female Reproductive Tract ◽  
Cryptic Female Choice ◽  
Trade Offs ◽  
Morphological Criteria

The field of post-copulatory sexual selection investigates how female and male adaptations have evolved to influence the fertilization of eggs while optimizing fitness during and after copulation, when females mate with multiple males. When females are polyandrous (one female mates with multiple males), they may optimize their mating rate and control the outcome of mating interactions to acquire direct and indirect benefits. Polyandry may also favor the evolution of male traits that offer an advantage in post-copulatory male-male sperm competition. Sperm competition occurs when the sperm, seminal fluid, and/or genitalia of one male directly impacts the outcome of fertilization success of a rival male. When a female mates with multiple males, she may use information from a number of traits to choose who will sire her offspring. This cryptic female choice (CFC) to bias paternity can be based on behavioral, physiological, and morphological criteria (e.g., copulatory courtship, volume and/or composition of seminal fluid, shape of grasping appendages). Because male fitness interests are rarely perfectly aligned with female fitness interests, sexual conflict over mating and fertilization commonly occur during copulatory and post-copulatory interactions. Post-copulatory interactions inherently involve close associations between female and male reproductive characteristics, which in many species potentially include sperm storage and sperm movement inside the female reproductive tract, and highlight the intricate coevolution between the sexes. This coevolution is also common in genital morphology. The great diversity of genitalia among species is attributed to sexual selection. The evolution of genital attributes that allow females to maintain reproductive autonomy over paternity via cryptic female choice or that prevent male manipulation and sexual control via sexually antagonistic coevolution have been well documented. Additionally, cases where genitalia evolve through intrasexual competition are well known. Another important area of study in post-copulatory sexual selection is the examination of trade-offs between investments in pre-copulatory and post-copulatory traits, since organisms have limited energetic resources to allocate to reproduction, and securing both mating and fertilization is essential for reproductive success.

scholarly journals Copulation, genital damage and early death in Callosobruchus maculatus

2006 ◽  
Vol 274 (1607) ◽  
pp. 247-252 ◽  
Author(s):  
Paul E Eady ◽  
Leticia Hamilton ◽  
Ruth E Lyons
Keyword(s):  
Reproductive Tract ◽  
Callosobruchus Maculatus ◽  
Seminal Fluid ◽  
Negative Impact ◽  
Early Death ◽  
Female Reproductive Tract ◽  
Mating Frequency ◽  
Trade Off ◽  
Female Longevity ◽  
Fluid Transfer

Antagonistic sexual coevolution stems from the notion that male and female interests over reproduction are in conflict. Such conflicts appear to be particularly obvious when male genital armature inflicts damage to the female reproductive tract resulting in reduced female longevity. However, studies of mating frequency, genital damage and female longevity are difficult to interpret because females not only sustain more genital damage, but also receive more seminal fluid when they engage in multiple copulations. Here, we attempt to disentangle the effects of genital damage and seminal fluid transfer on female longevity in the beetle Callosobruchus maculatus (Coleoptera: Bruchidae). Males copulating for the sixth time in succession inflicted greater levels of genital damage, but transferred smaller ejaculates in comparison with virgin males. The number of copulations performed by males was negatively related to female fecundity and positively related to female longevity, suggesting a trade-off between fecundity and longevity. However, inclusion of fecundity as a covariate revealed sperm and/or seminal fluid transfer to have a negative impact on female longevity above that caused by the fecundity–longevity trade-off. The consequences of multiple copulations on female longevity were examined. Females that mated twice laid more eggs and died sooner than those that mated once. However, incorporation of fecundity as a covariate into our statistical model removed the effect of female mating frequency on female longevity, indicating that double-mated females suffer greater mortality owing to the trade-off between fecundity and longevity. Males of this species are known to transfer very large ejaculates (up to 8% of their body weight), which may represent a significant nutritional benefit to females. However, the receipt of large ejaculates appears to carry costs. Thus, the interpretation of multiple mating experiments on female longevity and associated functional explanations of polyandry in this species are likely to be complex.

scholarly journals Inhibition of reverse transcriptases by seminalplasmin

1983 ◽  
Vol 209 (1) ◽  
pp. 183-188 ◽  
Author(s):  
E S P Reddy ◽  
M R Das ◽  
E P Reddy ◽  
P M Bhargava
Keyword(s):  
Dna Polymerase ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
Potent Inhibitor ◽  
Female Reproductive Tract ◽  
Dna Polymerase I ◽  
Avian Myeloblastosis Virus ◽  
Antibacterial Protein ◽  
Reverse Transcriptases ◽  
Polymerase I

Seminalplasmin, an antibacterial protein present in bovine seminal plasma, is shown to be a potent inhibitor of reverse transcriptases (RNA-dependent DNA nucleotidyltransferases). Seminalplasmin inhibits RNA-directed, hybrid-directed, and DNA-directed DNA-polymerizing activities of purified reverse transcriptase from avian myeloblastosis virus and from crude viral lysates of several retroviruses by binding to the enzyme, at least in the case of avian myeloblastosis virus. Seminalplasmin does not inhibit significantly DNA synthesis either by Escherichia coli DNA polymerase I, or a mammalian alpha-DNA polymerase. The presence of seminalplasmin in the seminal fluid could provide protection to the male and/or the female reproductive tract against retroviruses.

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

2012 ◽  
Vol 279 (1746) ◽  
pp. 4423-4432 ◽  
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.

scholarly journals Drosophila female reproductive tract gene expression reveals coordinated mating responses and rapidly evolving tissue-specific genes

2021 ◽  
Author(s):  
Caitlin E McDonough-Goldstein ◽  
Kirill Borziak ◽  
Scott Pitnick ◽  
Steve Dorus
Keyword(s):  
Gene Expression ◽  
Reproductive Tract ◽  
Expression Profiles ◽  
Female Reproductive Tract ◽  
Tissue Specific ◽  
Accelerated Evolution ◽  
Before And After ◽  
Complex Coordination ◽  
Extracellular Environment

Abstract Sexual reproduction in internally fertilizing species requires complex coordination between female and male reproductive systems and among the diverse tissues of the female reproductive tract (FRT). Here, we report a comprehensive, tissue-specific investigation of Drosophila melanogaster FRT gene expression before and after mating. We identified expression profiles that distinguished each tissue, including major differences between tissues with glandular or primarily non-glandular epithelium. All tissues were enriched for distinct sets of genes possessing secretion signals and exhibiting accelerated evolution, as might be expected for genes participating in molecular interactions between the sexes within the FRT extracellular environment. Despite robust transcriptional differences between tissues, post-mating responses were dominated by coordinated transient changes indicative of an integrated systems-level functional response. This comprehensive characterization of gene expression throughout the FRT identifies putative female contributions to post-copulatory events critical to reproduction and potentially reproductive isolation, as well as the putative targets of sexual selection and conflict.

scholarly journals proDA: Probabilistic Dropout Analysis for Identifying Differentially Abundant Proteins in Label-Free Mass Spectrometry

2020 ◽  
Author(s):  
Constantin Ahlmann-Eltze ◽  
Simon Anders
Keyword(s):  
Mass Spectrometry ◽  
Quantitative Proteomics ◽  
Statistical Power ◽  
Missing Values ◽  
Ad Hoc ◽  
Linear Models ◽  
Statistical Tests ◽  
High Sensitivity ◽  
Small Sample ◽  
Label Free

Abstract Protein mass spectrometry with label-free quantification (LFQ) is widely used for quantitative proteomics studies. Nevertheless, well-principled statistical inference procedures are still lacking, and most practitioners adopt methods from transcriptomics. These, however, cannot properly treat the principal complication of label-free proteomics, namely many non-randomly missing values. We present proDA, a method to perform statistical tests for differential abundance of proteins. It models missing values in an intensity-dependent probabilistic manner. proDA is based on linear models and thus suitable for complex experimental designs, and boosts statistical power for small sample sizes by using variance moderation. We show that the currently widely used methods based on ad hoc imputation schemes can report excessive false positives, and that proDA not only overcomes this serious issue but also offers high sensitivity. Thus, proDA fills a crucial gap in the toolbox of quantitative proteomics.

Sign in / Sign up

Export Citation Format

Share Document