scholarly journals Quantitative proteomics reveals rapid divergence in the postmating response of female reproductive tracts among sibling species

2020 ◽  
Vol 287 (1929) ◽  
pp. 20201030
Author(s):  
Erin L. McCullough ◽  
Caitlin E. McDonough ◽  
Scott Pitnick ◽  
Steve Dorus

Fertility depends, in part, on interactions between male and female reproductive proteins inside the female reproductive tract (FRT) that mediate postmating changes in female behaviour, morphology, and physiology. Coevolution between interacting proteins within species may drive reproductive incompatibilities between species, yet the mechanisms underlying postmating–prezygotic (PMPZ) isolating barriers remain poorly resolved. Here, we used quantitative proteomics in sibling Drosophila species to investigate the molecular composition of the FRT environment and its role in mediating species-specific postmating responses. We found that (i) FRT proteomes in D. simulans and D. mauritiana virgin females express unique combinations of secreted proteins and are enriched for distinct functional categories, (ii) mating induces substantial changes to the FRT proteome in D. mauritiana but not in D. simulans , and (iii) the D. simulans FRT proteome exhibits limited postmating changes irrespective of whether females mate with conspecific or heterospecific males, suggesting an active female role in mediating reproductive interactions. Comparisons with similar data in the closely related outgroup species D. melanogaster suggest that divergence is concentrated on the D. simulans lineage. Our study suggests that divergence in the FRT extracellular environment and postmating response contribute to previously described patterns of PMPZ isolation and the maintenance of species boundaries.

mBio ◽  
2019 ◽  
Vol 10 (2) ◽  
Author(s):  
Megan E. Spurgeon ◽  
Aayushi Uberoi ◽  
Stephanie M. McGregor ◽  
Tao Wei ◽  
Ella Ward-Shaw ◽  
...  

ABSTRACTPapillomaviruses exhibit species-specific tropism, thereby limiting understanding and research of several aspects of HPV infection and carcinogenesis. The discovery of a murine papillomavirus (MmuPV1) provides the opportunity to study papillomavirus infections in a tractable,in vivolaboratory model. MmuPV1 infects and causes disease in the cutaneous epithelium, as well as the mucosal epithelia of the oral cavity and anogenital tract. In this report, we describe a murine model of MmuPV1 infection and neoplastic disease in the female reproductive tracts of wild-type immunocompetent FVB mice. Low-grade dysplastic lesions developed in reproductive tracts of FVB mice infected with MmuPV1 for 4 months, and mice infected for 6 months developed significantly worse disease, including squamous cell carcinoma (SCC). We also tested the contribution of estrogen and/or UV radiation (UVR), two cofactors we previously identified as being involved in papillomavirus-mediated disease, to cervicovaginal disease. Similar to HPV16 transgenic mice, exogenous estrogen treatment induced high-grade precancerous lesions in the reproductive tracts of MmuPV1-infected mice by 4 months and together with MmuPV1 efficiently induced SCC by 6 months. UV radiation and exogenous estrogen cooperated to promote carcinogenesis in MmuPV1-infected mice. This murine infection model represents the first instance ofde novopapillomavirus-mediated carcinogenesis in the female reproductive tract of wild-type mice resulting from active virus infection and is also the first report of the female hormone estrogen contributing to this process. This model will provide an additional platform for fundamental studies on papillomavirus infection, cervicovaginal disease, and the role of cellular cofactors during papillomavirus-induced carcinogenesis.IMPORTANCETractable and efficient models of papillomavirus-induced pathogenesis are limited due to the strict species-specific and tissue-specific tropism of these viruses. Here, we report a novel preclinical murine model of papillomavirus-induced cervicovaginal disease in wild-type, immunocompetent mice using the recently discovered murine papillomavirus, MmuPV1. In this model, MmuPV1 establishes persistent viral infections in the mucosal epithelia of the female reproductive tract, a necessary component needed to accurately mimic HPV-mediated neoplastic disease in humans. Persistent MmuPV1 infections were able to induce progressive neoplastic disease and carcinogenesis, either alone or in combination with previously identified cofactors of papillomavirus-induced disease. This new model will provide a much-needed platform for basic and translational studies on both papillomavirus infection and associated disease in immunocompetent mice.


Author(s):  
Yasir H Ahmed-Braimah ◽  
Mariana F Wolfner ◽  
Andrew G Clark

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.


Author(s):  
Yasir H. Ahmed-Braimah ◽  
Mariana F. Wolfner ◽  
Andrew G. Clark

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.


Author(s):  
Caitlin E McDonough-Goldstein ◽  
Kirill Borziak ◽  
Scott Pitnick ◽  
Steve Dorus

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.


2006 ◽  
Vol 361 (1466) ◽  
pp. 261-268 ◽  
Author(s):  
Tami M Panhuis ◽  
Nathaniel L Clark ◽  
Willie J Swanson

Observations from different taxa, including plants, protozoa, insects and mammals, indicate that proteins involved in reproduction evolve rapidly. Several models of adaptive evolution have been proposed to explain this phenomenon, such as sexual conflict, sexual selection, self versus non-self recognition and pathogen resistance. Here we discuss the potential role of sexual conflict in the rapid evolution of reproductive genes in two different animal systems, abalone ( Haliotis ) and Drosophila . In abalone, we reveal how specific interacting sperm–egg proteins were identified and discuss this identification in the light of models for rapid protein evolution and speciation. For Drosophila , we describe the genomic approaches taken to identify male accessory gland proteins and female reproductive tract proteins. Patterns of protein evolution from both abalone and Drosophila support the predicted patterns of rapid protein evolution driven by sexual conflict. We stress however that other selective pressures may contribute to the rapid evolution that is observed. We conclude that the key to distinguishing between sexual conflict and other mechanisms of protein evolution will be an integration of genetic, experimental and theoretical data.


2020 ◽  
Vol 60 (3) ◽  
pp. 782-795 ◽  
Author(s):  
Teri J Orr ◽  
Virginia Hayssen

Synopsis Philosophical truths are hidden in Lewis Carroll’s nonsense poems, such as “The hunting of the snark.” When the poem is used as a scientific allegory, a snark stands for the pursuit of scientific truth, while a boojum is a spurious discovery. In the study of female biology, boojums have been the result of the use of cultural stereotypes to frame hypotheses and methodologies. Although female reproduction is key for the continuation of sexually reproducing species, not only have females been understudied in many regards, but also data have commonly been interpreted in the context of now-outdated social mores. Spurious discoveries, boojums, are the result. In this article, we highlight specific gaps in our knowledge of female reproductive biology and provide a jumping-off point for future research. We discuss the promise of emerging methodologies (e.g., micro-CT scanning, high-throughput sequencing, proteomics, big-data analysis, CRISPR-Cas9, and viral vector technology) that can yield insights into previously cryptic processes and features. For example, in mice, deoxyribonucleic acid sequencing via chromatin immunoprecipitation followed by sequencing is already unveiling how epigenetics lead to sex differences in brain development. Similarly, new explorations, including microbiome research, are rapidly debunking dogmas such as the notion of the “sterile womb.” Finally, we highlight how understanding female reproductive biology is well suited to the National Science Foundation’s big idea, “Predicting Rules of Life.” Studies of female reproductive biology will enable scholars to (1) traverse levels of biological organization from reproductive proteins at the molecular level, through anatomical details of the ovum and female reproductive tract, into physiological aspects of whole-organism performance, leading to behaviors associated with mating and maternal care, and eventually reaching population structure and ecology; (2) discover generalizable rules such as the co-evolution of maternal-offspring phenotypes in gestation and lactation; and (3) predict the impacts of changes to reproductive timing when the reliability of environmental cues becomes unpredictable. Studies in these key areas relative to female reproduction are sure to further our understanding across a range of diverse taxa.


1964 ◽  
Vol 46 (2) ◽  
pp. 217-229 ◽  
Author(s):  
E. S. E. Hafez

ABSTRACT The effects of ovariectomy and exogenous gonadal hormones on uterine enzymes have been examined for several laboratory mammals. Qualitative and quantitative measurement of uterine and placental enzymes involve histochemical, cytochemical and biochemical techniques. Enzymes have a wide and characteristic distribution in the female reproductive tract. The pattern of distribution varies showing peculiar species – specific differences in the anatomical localization of the enzyme and its relationship to the reproductive cycle. Progestational responses involve major endometrial growth and striking change in the enzyme system. The amount and biochemical composition of endometrial secretions vary with the stage of reproductive cycle. The placenta contains a complex enzyme system which has been classified into five major groups: hydrases, transferases, oxidoreductases, lyases and isomerases. The passage of many substances across the placental barrier is chemically regulated; the cytoplasm of barrier contains several enzymes and organelles. Carbonic anhydrase activity of the endometrium increases markedly during the luteal phase of the cycle and during pregnancy in the rabbit but not in some other species. Alkaline phosphatase is widely distributed in all types of placenta, in maximal concentration in species producing no fructose. Several hydrolytic enzymes are associated with the small cytoplasmic particles, the lysosomes, β-glucuronidase of the endometrial and glandular epithelia shows cyclical changes with the oestrus cycle. The activity of the succinic dehydrogenase may be an index of the functional status of the epithelial cells. Some enzymes e. g. acid and alkaline phosphatase, 5-nucleotidase and adenosinetriphosphate show a typical »secretion cycle«. The role of certain uterine enzymes in implantation of blastocyst deserves investigation.


2020 ◽  
Author(s):  
Martin D. Garlovsky ◽  
Caroline Evans ◽  
Mathew A. Rosenow ◽  
Timothy L. Karr ◽  
Rhonda R. Snook

ABSTRACTDespite holding a central role for fertilisation success, reproductive traits often show elevated rates of evolution and diversification. The rapid evolution of seminal fluid proteins (Sfps) within populations is predicted to cause mis-signalling between the male ejaculate and female reproductive tract between populations resulting in postmating prezygotic (PMPZ) isolation. Crosses between populations of Drosophila montana show PMPZ isolation in the form of reduced fertilisation success in both noncompetitive and competitive contexts. Here we test whether male ejaculate proteins deriving from either the accessory glands or the ejaculatory bulb differ between populations using liquid chromatography tandem mass spectrometry. We find more than 150 differentially abundant proteins between populations which may contribute to PMPZ isolation. These proteins include a number of proteases and peptidases, and several orthologs of D. melanogaster Sfps, all known to mediate fertilisation success and which mimic PMPZ isolation phenotypes. Males of one population typically produced greater quantities of Sfps and the strongest PMPZ isolation occurs in this direction. The accessory glands and ejaculatory bulb have different functions and the ejaculatory bulb contributes more to population differences than the accessory glands. Proteins with a secretory signal, but not Sfps, evolve faster than non-secretory proteins although the conservative criteria used to define Sfps may have impaired the ability to identify rapidly evolving proteins. We take advantage of quantitative proteomics data from three Drosophila species to determine shared and unique functional enrichments of Sfps that could be subject to selection between taxa and subsequently mediate PMPZ isolation. Our study provides the first high throughput quantitative proteomic evidence showing divergence of reproductive proteins implicated in the emergence of PMPZ isolation between populations.IMPACT SUMMARYIdentifying traits that prevent successful interbreeding is key to understanding early stages of the formation of new species, or speciation. Reproductive isolation arising prior to and during fertilisation frequently involves differences in how the sexes interact. In internally fertilising taxa, such interactions are mediated between the female reproductive tract where fertilisation occurs and the receipt of the ejaculate necessary for fertilisation. Because ejaculate proteins are at least partially responsible for these interactions, differences in male ejaculate protein composition could negatively impact fertilisation success, generating reproductive isolation. While the biological classes of ejaculate proteins are shared across all animal taxa, proteins that are secreted by males tend to show rapid evolution in gene expression and genetic sequence. Thus, reproductive proteins are suggested as prime targets facilitating reproductive isolation that arises after mating but before fertilisation (PostMating PreZygotic or PMPZ isolation). Most research on PMPZ isolation has focussed on differences between species for which it is not possible to determine the causative and temporal order of early speciation processes. Here, we test whether populations that exhibit few genetic differences but show strong PMPZ isolation also exhibit variation in ejaculate composition using quantitative high throughput proteomic analyses. We find a number of proteins are differentially abundant between populations including several known to impact fertilisation success in other species. We show that secreted proteins are evolving at an elevated rate, implicating their potential role in PMPZ isolation. We test divergence in ejaculate composition between species, finding a core set of functions that were conserved across species which last shared a common ancestor more than 40 million years ago along with species-specific investment. This work highlights the divergent evolution of reproductive proteins which may contribute to barriers between populations within a species early during speciation, extendable to similar analyses in other taxa in the future.


2018 ◽  
Author(s):  
Irem Sepil ◽  
Ben R Hopkins ◽  
Rebecca Dean ◽  
Marie-Laëtitia Thézénas ◽  
Philip D Charles ◽  
...  

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.


Author(s):  
Mai M. Said ◽  
Ramesh K. Nayak ◽  
Randall E. McCoy

Burgos and Wislocki described changes in the mucosa of the guinea pig uterus, cervix and vagina during the estrous cycle investigated by transmission electron microscopy. More recently, Moghissi and Reame reported the effects of progestational agents on the human female reproductive tract. They found drooping and shortening of cilia in norgestrel and norethindrone- treated endometria. To the best of our knowledge, no studies concerning the effects of mestranol and norethindrone given concurrently on the three-dimensional surface features on the uterine mucosa of the guinea pig have been reported. The purpose of this study was to determine the effect of mestranol and norethindrone on surface ultrastructure of guinea pig uterus by SEM.Seventy eight animals were used in this study. They were allocated into two groups. Group 1 (20 animals) was injected intramuscularly 0.1 ml vegetable oil and served as controls.


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