BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior

Male reproductive glands secrete signals into seminal fluid to facilitate reproductive success. In Drosophila melanogaster, these signals are generated by a variety of seminal peptides, many produced by the accessory glands (AGs). One epithelial cell type in the adult male AGs, the secondary cell (SC), grows selectively in response to bone morphogenetic protein (BMP) signaling. This signaling is involved in blocking the rapid remating of mated females, which contributes to the reproductive advantage of the first male to mate. In this paper, we show that SCs secrete exosomes, membrane-bound vesicles generated inside late endosomal multivesicular bodies (MVBs). After mating, exosomes fuse with sperm (as also seen in vitro for human prostate-derived exosomes and sperm) and interact with female reproductive tract epithelia. Exosome release was required to inhibit female remating behavior, suggesting that exosomes are downstream effectors of BMP signaling. Indeed, when BMP signaling was reduced in SCs, vesicles were still formed in MVBs but not secreted as exosomes. These results demonstrate a new function for the MVB–exosome pathway in the reproductive tract that appears to be conserved across evolution.

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Dynamic expression of bone morphogenetic protein 4 in reproductive organs of female mice

Reproduction ◽

10.1530/rep-10-0299 ◽

2011 ◽

Vol 142 (4) ◽

pp. 573-579 ◽

Cited By ~ 19

Author(s):

Pradeep S Tanwar ◽

James R McFarlane

Keyword(s):

Bone Morphogenetic Protein ◽

Granulosa Cells ◽

Reproductive Tract ◽

Follicular Development ◽

Reproductive Organs ◽

Bmp Signaling ◽

Female Reproductive Tract ◽

Specific Staining ◽

Primordial Follicles ◽

Morphogenetic Protein

Various members of the bone morphogenetic protein (BMP) family have been shown to regulate mammalian follicular development by affecting granulosa cell proliferation and steroidogenesis.In situhybridization studies have shown expression of BMPR1A, BMPR1B, and BMPR2 in the granulosa cells and oocyte of most of the follicles in the ovary, suggesting that these cells have the capacity to respond to BMP signaling. Although much is known about BMP4 signaling, its expression pattern in the female reproductive tract (FRT) is still unclear. The objective of the current study was to characterize the expression of BMP4 and its downstream target proteins (pSMAD1/5/8) in the FRT. In the ovary, BMP4 protein was detected in all the stages of follicular development. Staining for pSMAD1/5/8 was observed in granulosa cells and oocytes of all the stages of follicular development including primordial follicles, suggesting that these follicles are responsive to autocrine/paracrine BMP signaling. In the uterus, BMP4 and pSMAD1/5/8 staining was observed in all three compartments and strongest expression was observed during the estrus phase. BMP4- and pSMAD1/5/8-specific staining was also observed in oviductal epithelium. Different forms (apparent MW: 50, 35, and 15 kDa) of BMP4 were detected in mouse ovary by western blot analysis. In conclusion, these results have defined BMP4 and pSMAD1/5/8 protein expression in the mouse FRT and highlighted the importance of BMP4 in folliculogenesis.

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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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Seminal fluid protein divergence among populations exhibiting postmating prezygotic reproductive isolation

10.1101/2020.06.08.140020 ◽

2020 ◽

Author(s):

Martin D. Garlovsky ◽

Caroline Evans ◽

Mathew A. Rosenow ◽

Timothy L. Karr ◽

Rhonda R. Snook

Keyword(s):

Reproductive Isolation ◽

High Throughput ◽

Reproductive Tract ◽

Seminal Fluid ◽

Rapid Evolution ◽

Female Reproductive Tract ◽

Reproductive Proteins ◽

Fertilisation Success ◽

Accessory Glands ◽

Ejaculatory Bulb

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.

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Organoids of the female reproductive tract

Journal of Molecular Medicine ◽

10.1007/s00109-020-02028-0 ◽

2021 ◽

Vol 99 (4) ◽

pp. 531-553 ◽

Cited By ~ 1

Author(s):

Cindrilla Chumduri ◽

Margherita Y. Turco

Keyword(s):

Reproductive Tract ◽

Personalised Medicine ◽

Three Dimensional ◽

In Vitro Models ◽

Experimental Models ◽

Female Reproductive Tract ◽

Cellular Heterogeneity ◽

Dynamic Regulation ◽

Pregnancy And Childbirth

AbstractHealthy functioning of the female reproductive tract (FRT) depends on balanced and dynamic regulation by hormones during the menstrual cycle, pregnancy and childbirth. The mucosal epithelial lining of different regions of the FRT—ovaries, fallopian tubes, uterus, cervix and vagina—facilitates the selective transport of gametes and successful transfer of the zygote to the uterus where it implants and pregnancy takes place. It also prevents pathogen entry. Recent developments in three-dimensional (3D) organoid systems from the FRT now provide crucial experimental models that recapitulate the cellular heterogeneity and physiological, anatomical and functional properties of the organ in vitro. In this review, we summarise the state of the art on organoids generated from different regions of the FRT. We discuss the potential applications of these powerful in vitro models to study normal physiology, fertility, infections, diseases, drug discovery and personalised medicine.

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Post-Copulatory Sexual Selection

Evolutionary Biology ◽

10.1093/obo/9780199941728-0131 ◽

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.

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Copulation, genital damage and early death in Callosobruchus maculatus

Proceedings of The Royal Society B Biological Sciences ◽

10.1098/rspb.2006.3710 ◽

2006 ◽

Vol 274 (1607) ◽

pp. 247-252 ◽

Cited By ~ 58

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.

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Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.611301 ◽

2021 ◽

Vol 8 ◽

Author(s):

Eduardo G. Aisen ◽

Wilfredo Huanca López ◽

Manuel G. Pérez Durand ◽

Edita Torres Mamani ◽

Juan C. Villanueva Mori ◽

...

Keyword(s):

Seminal Plasma ◽

Artificial Insemination ◽

Reproductive Tract ◽

Vas Deferens ◽

Female Reproductive Tract ◽

Low Fertility ◽

Sperm Cells ◽

South American Camelids ◽

Thawing Process

The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge.

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Development of parthenogenetic and fertilized mouse embryos in the uterus and in extra-uterine sites

Development ◽

10.1242/dev.34.2.387 ◽

1975 ◽

Vol 34 (2) ◽

pp. 387-405

Author(s):

S. A. Iles ◽

M. W. McBurney ◽

S. R. Bramwell ◽

Z. A. Deussen ◽

C. F. Graham

Keyword(s):

Reproductive Tract ◽

Neural Tissue ◽

Cell Types ◽

Mouse Embryos ◽

Female Reproductive Tract ◽

Embryonic Cells ◽

Haploid Embryos ◽

Keratinized Epithelium ◽

Mouse Eggs

Mouse eggs were activated with hyaluronidase in vitro and subsequently transferred to the oviduct. In the female reproductive tract they formed morulae and blastocysts which died soon after implantation. Haploid blastocysts were transferred beneath the kidney capsule and here some formed disorganized egg-cylinder structures in a week. Morulae and blastocysts from haploid and diploid parthenogenones were also transferred beneath the testis capsule. Two to four months later the growths which had formed were sectioned. They contained neural tissue, pigment, keratinized epithelium, glandular epithelium, ciliated epithelium, cartilage, bone, muscle, adipose tissue, and haemopoietic tissue. The range of cell types was similar to that produced by fertilized control blastocysts except that the parthenogenones did not form identifiable yolk-sac carcinoma or embryonal carcinomacells. The growths from haploid and diploid parthenogenones in the testis were stained with Feulgen and their DNA content measured. Growths from diploid embryos contained the normal diploid amount of DNA while growths from haploid embryos contained less than this amount. Cell cultures were prepared from the growths. The cells which were investigated contained no Y chromosome, suggesting that they were derived from the embryonic cells rather than the cells of the male host. These cells contained a near diploid chromosome number, although some of them were originally derived from haploid embryos.

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CatSperζ regulates the structural continuity of sperm Ca2+ signaling domains and is required for normal fertility

eLife ◽

10.7554/elife.23082 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 54

Author(s):

Jean-Ju Chung ◽

Kiyoshi Miki ◽

Doory Kim ◽

Sang-Hee Shim ◽

Huanan F Shi ◽

...

Keyword(s):

Reproductive Tract ◽

Female Reproductive Tract ◽

Targeted Disruption ◽

Vitro Fertilization ◽

Male Subfertility ◽

Epididymal Spermatozoa ◽

Mammalian Female ◽

Signaling Domains

We report that the Gm7068 (CatSpere) and Tex40 (CatSperz) genes encode novel subunits of a 9-subunit CatSper ion channel complex. Targeted disruption of CatSperz reduces CatSper current and sperm rheotactic efficiency in mice, resulting in severe male subfertility. Normally distributed in linear quadrilateral nanodomains along the flagellum, the complex lacking CatSperζ is disrupted at ~0.8 μm intervals along the flagellum. This disruption renders the proximal flagellum inflexible and alters the 3D flagellar envelope, thus preventing sperm from reorienting against fluid flow in vitro and efficiently migrating in vivo. Ejaculated CatSperz-null sperm cells retrieved from the mated female uterus partially rescue in vitro fertilization (IVF) that failed with epididymal spermatozoa alone. Human CatSperε is quadrilaterally arranged along the flagella, similar to the CatSper complex in mouse sperm. We speculate that the newly identified CatSperζ subunit is a late evolutionary adaptation to maximize fertilization inside the mammalian female reproductive tract.

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301 THE EFFECT OF EQUINE FOLICULLAR FLUID ON IN VITRO INDUCTION OF THE ACROSOME REACTION IN FROZEN - THAWED STALLION SPERMATOZOA

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab301 ◽

2010 ◽

Vol 22 (1) ◽

pp. 307

Author(s):

D. S. Silva ◽

P. Rodriguez ◽

N. S. Arruda ◽

R. Rodrigues ◽

J. L. Rodrigues

Keyword(s):

Contact Area ◽

Follicular Fluid ◽

Acrosome Reaction ◽

Reproductive Tract ◽

Female Reproductive Tract ◽

Heparin Group ◽

Fluorescent Stain ◽

In Vitro Induction

The capacitation process occurs in vivo upon exposure of the spermatozoa through the female reproductive tract, but can be induced in vitro in the presence of several compounds. This study was conducted to assess the effect of heparin or equine follicular fluid on hyperactivated motility and in vitro induction acrosome reaction swim-up method with frozen-thawed stallion semen. Two hundred microliters of frozen-thawed equine semen was placed in a tube (45°C) to increase contact area and incubated at 37°C for 1 h. After incubation 800 μL of the supernatant was collected by centrifugation (500 × g, 10 min) to collect spermatozoa. The resulting pellet was resuspended in capacitation medium Fert-TALP supplemented with 5.0 μg mL-1 heparin or 100% follicular fluid and incubated for different times (1, 2, 3, 4, and 5 h) at 37°C. After incubation the hyperactivated motility and acrosome-reacted spermatozoa were evaluated. Hoechst stain was used to differentiate live and dead spermatozoa, and chlortetracycline (CTC) fluorescent stain was used to assess the capacitation response of sperm; data were analyzed by ANOVA. The effect of equine follicular fluid resulted in improved percentage of spermatozoa with acrosome reaction at all times of incubation (60, 63, 57, 52, and 58%) but immediately after 3 h of incubation, the hyperactivated motility decreased in heparin group and follicular fluid (42 and 30%, respectively).

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