Step In Reproduction Research Articles

Identification of Intercellular Crosstalk between Decidual Cells and Niche Cells in Mice.

Decidualization is a crucial step for human reproduction, which is a prerequisite for embryo implantation, placentation and pregnancy maintenance. Despite rapid advances over recent years, the molecular mechanism underlying decidualization remains poorly understood. Here, we used the mouse as an animal model and generated a single-cell transcriptomic atlas of a mouse uterus during decidualization. By analyzing the undecidualized inter-implantation site of the uterus as a control, we were able to identify global gene expression changes associated with decidualization in each cell type. Additionally, we identified intercellular crosstalk between decidual cells and niche cells, including immune cells, endothelial cells and trophoblast cells. Our data provide a valuable resource for deciphering the molecular mechanism underlying decidualization.

Proteomics based drug repositioning applied to improve in vitro fertilization implantation: an artificial intelligence model

ABSTRACT Embryo implantation is one of the most inefficient steps in assisted reproduction, so the identifying drugs with a potential clinical application to improve it has a strong interest. This work applies artificial intelligence and systems biology-based mathematical modeling strategies to unveil potential treatments by computationally analyzing and integrating available molecular and clinical data from patients. The mathematical models of embryo implantation computationally generated here simulate the molecular networks underneath this biological process. Once generated, these models were analyzed in order to identify potential repositioned drugs (drugs already used for other indications) able to improve embryo implantation by modulating the molecular pathways involved. Interestingly, the repositioning analysis has identified drugs considering two endpoints: (1) drugs able to modulate the activity of proteins whose role in embryo implantation is already bibliographically acknowledged, and (2) drugs that modulate key proteins in embryo implantation previously predicted through a mechanistic analysis of the mathematical models. This second approach increases the scope open for examination and potential novelty of the repositioning strategy. As a result, a list of 23 drug candidates to improve embryo implantation after IVF was identified by the mathematical models. This list includes many of the compounds already tested for this purpose, which reinforces the predictive capacity of our approach, together with novel repositioned candidates (e.g., Infliximab, Polaprezinc, and Amrinone). In conclusion, the present study exploits existing molecular and clinical information to offer new hypotheses regarding molecular mechanisms in embryo implantation and therapeutic candidates to improve it. This information will be very useful to guide future research. Abbreviations: IVF: in vitro fertilization; EI: Embryo implantation; TPMS: Therapeutic Performance Mapping System; MM: mathematical models; ANN: Artificial Neuronal Networks; TNFα: tumour necrosis factor factor-alpha; HSPs: heat shock proteins; VEGF: vascular endothelial growth factor; PPARA: peroxisome proliferator activated receptor-α PXR: pregnane X receptor; TTR: transthyretin; BED: Biological Effectors Database; MLP: multilayer perceptron

Cell-Cell Communication at the Embryo Implantation Site of Mouse Uterus Revealed by Single-Cell Analysis.

As a crucial step for human reproduction, embryo implantation is a low-efficiency process. Despite rapid advances in recent years, the molecular mechanism underlying embryo implantation remains poorly understood. Here, we used the mouse as an animal model and generated a single-cell transcriptomic atlas of embryo implantation sites. By analyzing inter-implantation sites of the uterus as control, we were able to identify global gene expression changes associated with embryo implantation in each cell type. Additionally, we predicted signaling interactions between uterine luminal epithelial cells and mural trophectoderm of blastocysts, which represent the key mechanism of embryo implantation. We also predicted signaling interactions between uterine epithelial-stromal crosstalk at implantation sites, which are crucial for post-implantation development. Our data provide a valuable resource for deciphering the molecular mechanism underlying embryo implantation.

Back to the future: optimised microwell culture of individual human preimplantation stage embryos.

Although in vitro culture of human embryos is a crucial step in assisted reproduction, the lack of focused research hampers worldwide standardisation and consistent outcomes. Only 1.2% of research papers published in five leading journals in human reproduction in 2019 focused on in vitro culture conditions, creating the impression that the optimisation process has approached its limits. On the other hand, in vitro culture of mammalian embryos is based on old principles, while there is no consensus on basic issues as density, time, medium change, gas atmosphere and small technical details including the way of drop preparation. This opinion paper aims to highlight and analyse the slow advancement in this field and stimulate research for simple and affordable solutions to meet the current requirements. A possible way for advancement is discussed in detail. Selection of embryos with the highest developmental competence requires individual culture and modification of the widely used "drop under oil" approach. Current use of three-dimensional surfaces instead of large flat bottoms is restricted to time-lapse systems, but these wells are designed for optical clarity, not for the needs of embryos. The size and shape of the original microwells (Well of the Well; WOW) offer a practical and straightforward solution to combine the benefits of communal and individual incubation and improve the overall quality of cultured embryos.

A Morphological and Molecular Analysis of a Bloom of the Filamentous Green Alga Pithophora

Filamentous representatives of Cladophorales (Chlorophyta) are major contributors to algal biomass of littoral communities. In the present study, community analysis of a reported bloom in the Ogeechee River in Georgia provided an opportunity to combine morphological and genetic analyses with ecological information related to an understudied nuisance alga. A polyphasic approach of incorporating genotypic and phenotypic methods led to the identification of the algal community as Pithophora roettleri (Roth) Wittrock. Morphological analysis showed a monospecific community based on the average length and diameter of the heterosporous intercalary and terminal akinetes, along with the diameter of the principal filaments. Single-gene and concatenated-gene phylogenetic analyses of the LSU (28S rRNA) and SSU (18S rRNA) markers further confirmed this species identification. In this study, we conducted a morphological treatment of P. roettleri, produced 17 novel gene sequences, and produced a new, schematic diagram illustrating the four steps of the asexual reproduction of an intercalary akinete. Morphological characteristics, like the position and shape of akinetes documented here, and the availability of genetic sequences can improve identification and further ecological understanding of filamentous green algae. Flowing mats of P. roettleri, like those observed in this study, can lower light availability for other biota and structurally alter the habitat.

Paving the Way for Fertilization: The Role of the Transmitting Tract.

Angiosperm reproduction relies on the precise growth of the pollen tube through different pistil tissues carrying two sperm cells into the ovules’ embryo sac, where they fuse with the egg and the central cell to accomplish double fertilization and ultimately initiate seed development. A network of intrinsic and tightly regulated communication and signaling cascades, which mediate continuous interactions between the pollen tube and the sporophytic and gametophytic female tissues, ensures the fast and meticulous growth of pollen tubes along the pistil, until it reaches the ovule embryo sac. Most of the pollen tube growth occurs in a specialized tissue—the transmitting tract—connecting the stigma, the style, and the ovary. This tissue is composed of highly secretory cells responsible for producing an extensive extracellular matrix. This multifaceted matrix is proposed to support and provide nutrition and adhesion for pollen tube growth and guidance. Insights pertaining to the mechanisms that underlie these processes remain sparse due to the difficulty of accessing and manipulating the female sporophytic tissues enclosed in the pistil. Here, we summarize the current knowledge on this key step of reproduction in flowering plants with special emphasis on the female transmitting tract tissue.

Synaptic communication mediates the assembly of a self-organizing circuit that controls reproduction.

Migration of gonadotropin-releasing hormone (GnRH) neurons from their birthplace in the nasal placode to their hypothalamic destination is critical for vertebrate reproduction and species persistence. While their migration mode as individual GnRH neurons has been extensively studied, the role of GnRH-GnRH cell communication during migration remains largely unexplored. Here, we show in awake zebrafish larvae that migrating GnRH neurons pause at the nasal-forebrain junction and form clusters that act as interhemisphere neuronal ensembles. Within the ensembles, GnRH neurons create an isolated, spontaneously active circuit that is internally wired through monosynaptic glutamatergic synapses into which newborn GnRH neurons integrate before entering the brain. This initial phase of integration drives a phenotypic switch, which is essential for GnRH neurons to properly migrate toward their hypothalamic destination. Together, these experiments reveal a critical step for reproduction, which depends on synaptic communication between migrating GnRH neurons.

Mortality risk promotes cooperation of wasps when paralysing hosts

The role of risk in behavioural choices leading to cooperation in attacking a prey or a host has proven to be difficult to assess in most systems because risk is difficult to quantify. For parasitoid wasps, paralysing a host is a key step for successful parasitism and reproduction, but wasps risk death or injury because of host defences and may require group action, which provides a model system to decipher the mechanism driving cooperation. We explored scenarios where wasps behave as either cooperators or free-riding defectors by using the parasitoid Sclerodermus guani and its host Monochamus alternatus. First, when one or two wasps paralysed a single host, the mortality of both increased with increasing host size but was significantly lower when there were two wasps. Second, fitness analysis showed that whether a wasp was involved in paralysing a host did not affect the number of eggs laid. Finally, when two wasps paralysed single large hosts, higher mortality risks led to more cooperation, whereas lower risk led to more frequent free riding. Our study implies that risk is involved in mediating the level of cooperation and provides an experimental demonstration of risk-driven cooperation of parasitoids.

The role of reproductive loss on flock performance: a comparison of nine industry flocks.

The reproductive performance of a sheep flock is dependent on a multitude of complex interacting factors. Attaining optimal flock performance requires information about how the reproductive steps are linked and relate to readily available measurements of the state of the flock. The goal was to use data from nine commercial flocks (greater than 300,000 records) to investigate and model the key reproductive steps affecting flock reproductive performance. We also developed a maximum-likelihood based methodology to predict flock ovulation rate based on measurements of the number of fetuses at mid-pregnancy (detected by ultrasound-scanning). The model was used to determine how changes in premating liveweight, age, predicted ovulation rate, number of fetuses at mid-pregnancy, lamb survival and lamb growth rate affect the total lamb liveweight at weaning per ewe exposed to the ram in each flock. The data from the commercial flocks were also used to investigate the role of ewe age and premating liveweight on each reproductive step. Sensitivity analyses were conducted to identify the key reproductive steps affecting flock reproductive performance, with a focus on understanding how these steps vary between flocks. The elasticity for embryo survival was 60% of that for lamb survival for these flocks and the elasticities for ovulation rate were highly variable between flocks (0.16 to 0.50 for mature ewes). This indicates that ovulation rate was near-optimal for some flocks, whereas there was potential to significantly improve flock performance in suboptimal flocks. The elasticity for ewe premating liveweight was highly variable between flocks (−0.03 to 0.84 for mature ewes and −0.18 to 1.39 for ewe lambs), indicating that premating liveweight ranged from optimal to suboptimal between flocks. For these suboptimal farms, the opportunity exists to increase flock performance through improved management of ewe premating liveweight. Reproductive loss was significantly greater in ewe lambs than mature ewes, although the difference is dependent on the stage of reproduction and flock. Predicted ovulation rate was 25% lower for ewe lambs and there was a 30% relative decrease in the predicted embryo survival probability from ovulation to scanning for ewe lambs. There was a 10% relative decrease in lamb survival probability from birth to weaning for ewe lambs and lamb growth rate was 25% lower for ewe lambs.

The histone demethylase KDM2B regulates human primordial germ cell-like cells specification

Germline specification is a fundamental step for human reproduction and this biological phenomenon possesses technical challenges to study in vivo as it occurs immediately after blastocyst implantation. The establishment of in vitro human primordial germ cell-like cells (hPGCLCs) induction system allows sophisticated characterization of human primordial germ cells (hPGCs) development. However, the underlying molecular mechanisms of hPGCLC specification are not fully elucidated. Here, we observed particularly high expression of the histone demethylase KDM2B in male fetal germ cells (FGCs) but not in male somatic cells. Besides, KDM2B shared similar expression pattern with hPGC marker genes in hPGCLCs, suggesting an important role of KDM2B in germ cell development. Although deletion of KDM2B had no significant effects on human embryonic stem cell (hESC)'s pluripotency, loss of KDM2B dramatically impaired hPGCLCs differentiation whereas ectopically expressed KDM2B could efficiently rescue such defect, indicating this defect was due to KDM2B's loss in hPGCLC specification. Mechanistically, as revealed by the transcriptional profiling, KDM2B suppressed the expression of somatic genes thus inhibited somatic differentiation during hPGCLC specification. These data collectively indicate that KDM2B is an indispensable epigenetic regulator for hPGCLC specification, shedding lights on how epigenetic regulations orchestrate transcriptional events in hPGC development for future investigation.

Sugar and Hormone Dynamics and the Expression Profiles of SUT/SUC and SWEET Sweet Sugar Transporters during Flower Development in Petunia axillaris.

Flowering is the first committed step of plant sexual reproduction. While the developing flower is a strong sink requiring large quantity of sugars from photosynthetic source tissues, this process is under-temper-spatially controlled via hormone signaling pathway and nutrient availability. Sugar transporters SUT/SUC and SWEET mediate sugars movement across membranes and play a significant role in various physiological processes, including reproductive organ development. In Petunia axillaris, a model ornamental plant, 5 SUT/SUC and 36 SWEET genes are identified in the current version of the genome. Analysis of their gene structure and chromosomal locations reveal that SWEET family is moderately expanded. Most of the transporter genes are abundantly expressed in the flower than in other organs. During the five flower developmental stages, transcript levels of PaSUT1, PaSUT3, PaSWEET13c, PaSWEET9a, PaSWEET1d, PaSWEET5a and PaSWEET14a increase with the maturation of the flower and reach their maximum in the fully open flowers. PaSWEET9c, the nectar-specific PhNEC1 orthologous, is expressed in matured and fully opened flowers. Moreover, determination of sugar concentrations and phytohormone dynamics in flowers at the five developmental stages shows that glucose is the predominant form of sugar in young flowers at the early stage but depletes at the later stage, whereas sucrose accumulates only in maturated flowers prior to the corolla opening. On the other hand, GA3 content and to a less extent IAA and zeatin decreases with the flower development; however, JA, SA and ABA display a remarkable peak at mid- or later flower developmental stage.

Dynamically swarm shared mutation based bacterial foraging

In this paper, dynamically swarm shared mutation based Bacterial Foraging (DSSBFO) is proposed to optimize multidimensional, unimodal and multimodal functions. In BFO, due to fixed step size it requires more computational cost to get optimum solution with better accuracy. Chemotaxis and reproduction step of BFO are not sufficient for an effective search. So in this paper, the authors propose dynamic step size in BFO to achieve optimum solution with better accuracy with minimum cost. The dynamic step size i.e. mutation is achieved by modifying the position equation of GLBestPSO and momentum factor (mc) of SSMPSO used in modified equation to bring the bacteria in search space and not to cross the boundary of search space. The eight standard benchmark functions are used to prove the performance of DSSBFO in terms of precision and cost. DSSBFO performs well as compared to BFO and BSO (BFO hybridized with PSO) alogrithms interms of quality solution with faster convergence.

An intelligent algorithm based on evolutionary strategy and clustering algorithm for Lamb wave defect location

Imaging algorithms for visualization of defects play a significant role in Lamb wave–based research of nondestructive testing and structural health monitoring. In classical algorithms, the position or distribution of defects is located by mapping the amplitude or phase information of signals from the time domain to every discrete spatial grid of the structure. It is time-consuming. In this study, the diversity, statistical, and fuzzy characteristics of the elliptic imaging algorithm are analyzed first; then, an intelligent defect location algorithm is proposed based on the evolutionary strategy and the K-means algorithm. The position of defects can be identified by observing the distribution of individuals. There are six parts in the proposed algorithm, including the data structure design, adaptive population screening, adaptive population reproduction, diversity maintenance mechanism, and cutoff criterion. Considering the statistical and fuzzy characteristics in the detection, several specific input parameters are defined in our algorithm, such as the distance-dependent screening threshold, path-dependent residual vector, and path-independent residual. To maintain the diversity of individuals in the analysis, we have made two adjustments to the evolutionary strategy: one is to optimize the population screening and reproduction steps with the K-means algorithm, and the other is to add a diversity maintenance method into the evolutionary strategy. The effectiveness of the proposed intelligent defect location algorithm is verified by numerical simulations and experiments. Numerical studies indicate that the proposed algorithm has a reliable performance in the detection of defects with different shapes and sizes. In the experimental research, we demonstrate that the efficiency of the proposed algorithm is about 200 times faster than the elliptic imaging algorithm. And the optimum parameter setting of the algorithm is investigated by analyzing the influence of parameter setting on the detection.

다색 협힐 재현을 위한 제작 기법 연구 - 일본 정창원 소장 협힐을 중심으로

The purpose of this study is to provide the basic data for reproducing the principles of the manufacturing method of the Clamp resist dyeing at that time by analyzing the artifacts of the Clamp resist dyeing in Shosoin and producing Wooden Printing Block and dyeing the template. To this end, director of Shosoin found clues for the production of the Clamp resist dyeing in Clamp resist dyeing pattern and actually produced and solved many of the questions arising from verifying each process. In particular, the method of manufacturing the Wooden Printing Block, the method of fixing the Wooden Printing Block, the method of sandwiching the fabric between the Wooden Printing Block, and the method of injecting dyeing solution were studied. Through this, it is meaningful to provide basic data of each dyeing step for reproduction of Clamp resist dyeing, and it is considered that continuous reproduction research will be required by selecting various materials and patterns in the future. Furthermore, it is expected that a variety of Clamp resist dyeing crafts will be developed by grafting with modern technology based on the reproduced dyeing technology.

Reproduction Function in Male Patients With Bardet Biedl Syndrome.

PurposeBardet-Biedl syndrome (BBS) is a ciliopathy with a wide spectrum of symptoms due to primary cilia dysfunction, including genitourinary developmental anomalies as well as impaired reproduction, particularly in males. Primary cilia are known to be required at the following steps of reproduction function: (i) genitourinary organogenesis, (ii) in fetal firing of hypothalamo-pituitary axe, (iii) sperm flagellum structure, and (iv) first zygotic mitosis conducted by proximal sperm centriole. BBS phenotype is not fully understood.MethodsThis study explored all steps of reproduction in 11 French male patients with identified BBS mutations.ResultsBBS patients frequently presented with genitourinary malformations, such as cryptorchidism (5/11), short scrotum (5/8), and micropenis (5/8), but unexpectedly, with normal testis size (7/8). Ultrasonography highlighted epididymal cysts or agenesis of one seminal vesicle in some cases. Sexual hormones levels were normal in all patients except one. Sperm numeration was normal in 8 out of the 10 obtained samples. Five to 45% of sperm presented a progressive motility. Electron microscopy analysis of spermatozoa did not reveal any homogeneous abnormality. Moreover, a psychological approach pointed to a decreased self-confidence linked to blindness and obesity explaining why so few BBS patients express a child wish.ConclusionsPrimary cilia dysfunction in BBS impacts the embryology of the male genital tract, especially epididymis, penis, and scrotum through an insufficient fetal androgen production. However, in adults, sperm structure does not seem to be impacted. These results should be confirmed in a greater BBS patient cohort, focusing on fertility.

Recent progress in bovine in vitro-derived embryo cryotolerance: Impact of in vitro culture systems, advances in cryopreservation and future considerations.

Cryopreservation of in vitro-derived bovine embryos is a crucial step for the widespread reproduction and conservation of valuable high-merit animals. Given the current popularity of bovine in vitro embryo production (IVP), there is a demand for a highly efficient ultra-low temperature storage method in order to maximize donor ovum pickup (OPU) turn-over, recipient availability/utilization and domestic/overseas commercial trading opportunities. However, IVP bovine embryos are still very sensitive to chilling and cryopreservation, and despite recent progress, a convenient (simple and robust) protocol has not yet been developed. At the moment, there are two methods for bovine IVP embryo cryopreservation: slow programmable freezing and vitrification. Both of the aforementioned techniques have pros and cons. While controlled-rate slow cooling can easily be adapted for direct transfer (DT), ice crystal formation remains an issue. On the other hand, vitrification solved this problem but the possibility of successful DT commercial incorporation remains to be determined. Moreover, simplification of the vitrification protocol (including warming) through the use of an in-straw dilution without the use of a microscope is a prerequisite for its use under farm conditions. This review summarizes the bovine IVP embryo cryopreservation achievements, strengths and limitations of both freezing systems and prospective improvements to enhance cryosurvival, as well as perspectives on future directions of this assisted reproductive technology.

The human factor: does the operator performing the embryo transfer significantly impact the cycle outcome?

STUDY QUESTIONIs Ongoing Pregnancy Rate (OPR) operator-dependent, and can experience improve embryo transfer efficiency?SUMMARY ANSWEROPR is influenced by the operators who perform the embryo transfer (ET), and experience does not assure proficiency for everyone.WHAT IS KNOWN ALREADYET remains the critical step in assisted reproduction. Although many other factors such as embryo quality and uterine receptivity impact embryo implantation, the proper ET technique is clearly an operator-dependent variable and as such it should be objectively standardized.STUDY DESIGN, SIZE, DURATIONRetrospective comparative analysis including all fresh ETs performed between January 1996 and December 2016 at the Humanitas Fertility Center after IVF—ICSI cycles.PARTICIPANTS/MATERIALS, SETTING, METHODSIVF/ICSI fresh ETs performed by 32 operators, 19 824 cycles in all, were analyzed. All transfers consisting of freehand insertion of a preloaded soft catheter into the uterine cavity under transabdominal ultrasound guidance were considered. Two different statistical analyses were performed. First, a logistic regression model with a random intercept for the operator was used to estimate the heterogeneity of the rate of success among operators, accounting for woman age, FSH, number of oocytes retrieved, fertilization rate, year of the procedure, number and stage of transferred embryos and operator’s experience. Second, the relationship between experience and pregnancy rate was estimated separately for each operator by logistic regression, and operator-specific results were combined and compared in a random-effects meta-analysis. In both analyses, the operator’s experience at time t was measured in terms of number of embryo transfers performed before t.MAIN RESULTS AND THE ROLE OF CHANCEThe heterogeneity among operators was highly significant (P value <0.001) and explained 44.5% of the total variability. The odds ratio of success of the worst operator in respect to the mean was equal to 0.84. For the best operator, the odds ratio of success was equal to 1.13 in respect to the mean. Based on the meta-analysis of the relationship between operator’s experience and success rate, it resulted that, on average, the operators’ performance did not improve with additional transfers.LIMITATIONS, REASONS FOR CAUTIONAt our center, operators become independent for ET’s after performing between 30 and 50 transfers under supervision. It is also possible that other relevant factors, such as embryologists on duty for the ET, have not been included in the present analysis and this may represent a potential bias. Among these, it should be mentioned that the embryologists on duty for the ET were not taken into consideration.WIDER IMPLICATIONS OF THE FINDINGSContinued performance analysis and the use of a digital simulator could help operators to test their expertise over time and either correct poor performance or avoid doing transfers.STUDY FUNDING/COMPETING INTEREST(S)None.TRIAL REGISTRATION NUMBERNCT03561129.

The effect of reproductive loss on the performance of a research flock.

The reproductive performance of a sheep flock is dependent on a multitude of complex interacting factors. Achieving optimal flock performance requires knowledge of the reproductive steps and how these are linked and related to available measurements of the state and performance of the flock. The goal was to use previously collected data from a research flock that had undergone selection for fecundity (11,369 lambing records), to model the key reproductive steps affecting flock reproductive performance. The model was used to investigate how changes in liveweight, age, ovulation rate, number of fetuses at midpregnancy, number of lambs born, and birthweight affect the number of lambs weaned and the weaning weight of each lamb in this flock. The data available from the research flock were used to parameterize models of each reproductive step and assess the role of ewe age and premating liveweight on each reproductive step. These models were then linked together as a simulation tool to assess the role of different parameters on flock reproductive performance, which was defined as the total weight of lambs weaned per ewe exposed to the ram. Flock elasticities were calculated that characterize the relative importance of the effect of average premating ewe liveweight (0.81), average ovulation rate (0.33), variance in ovulation rate (-0.095), embryo survival (0.72), lamb survival (1.03), conception failure (0.35), and average ewe age (0.056) on the total kilograms of lamb liveweight at weaning per ewe exposed to the ram. The largest elasticity for lamb survival indicated that a 1% increase in lamb survival is expected to have a 1.03% increase in the total kilograms of lamb liveweight at weaning per ewe exposed to the ram in this flock. Assuming similar costs, interventions to increase lamb survival for this flock will provide the largest increase in the total kilograms of lamb liveweight at weaning per ewe exposed to the ram, which is a key metric of flock performance.

Workflow to Characterize Mutants with Reproductive Defects.

Reverse genetics approaches for characterizing phenotypes of mutants in a gene of interest (GOI) require thorough genotyping and phenotypic analysis. However, special challenges are encountered when a GOI is expressed in reproductive tissues: a variety of assays are required to characterize the phenotype and a mutant may show sporophytic and/or gametophytic defects in male and/or female reproductive tissues, which are structurally and functionally intertwined. Here, we present a streamlined workflow to characterize mutants with reproductive defects, primarily using Arabidopsis as a model, which can also be adapted to characterize mutants in other flowering plants. Procedures described here can be used to distinguish different kinds of reproductive defects and pinpoint the defective reproductive step(s) in a mutant. Although our procedures emphasize the characterization of mutants with male reproductive defects, they can nevertheless be used to identify female reproductive defects, as those defects could manifest alongside, and sometimes require, male reproductive tissues.

How pirate phage interferes with helper phage: Comparison of the two distinct strategies

Pirate phages use the structural proteins encoded by unrelated helper phages to propagate. The best-studied example is the pirate P4 and helper P2 of coliphages, and it has been known that the Staphylococcus aureus pathogenicity islands (SaPIs) that can encode virulence factors act as pirate phages, too. When alone in the host, the pirate phages act as a prophage, but when the helper phage gene is also in the same host cell, the pirate phage has ability to exploit the helper phages structural proteins to produce pirate phage particles and spread, interfering with the helper phage production. The known helper phages in these systems are temperate phages. Interestingly, the interference of the pirate phage to the helper phage occurs in a different manner between the SaPI-helper system and the P4-P2 system. SaPIs cannot lyse a helper lysogen upon infection, while when a helper phage lyse a SaPI lysogen, most of the phage particles produced are the SaPI particles. On the contrary, in the P4-P2 system, a pirate phage P4 can lyse a helper P2 lysogen to produce mostly the P4 particles, while when P2 phage lyses a P4 lysogen, most of the produced phages are the P2 particles. Here, the consequences of these different strategies in the pirate and helper phage spreading among uninfected host is analyzed by using mathematical models. It is found that SaPI’s strategy interferes with the helper phage spreading significantly more than the P4’s strategy, because SaPI interferes with the helper phage’s main reproduction step, while P4 interferes only by forcing the helper lysogens to lyse. However, the interference is found to be weaker in the spatially structured environment than in the well-mixed environment. This is because, in the spatial setting, the system tends to self-organize so that the helper phages take over the front of propagation due to the need of helper phage for the pirate phage spreading.