Embryo Survival Research Articles

Nanoparticles Combining Host‐Directed Therapeutics and Antibiotics to Boost Bacterial Killing and Overall Survival of Zebrafish Embryos Infected with Mycobacterium Marinum

AbstractThe bacterial pathogen, Mycobacterium tuberculosis (Mtb), remains a leading cause of global morbidity and mortality. Due to the complexity of the infection, the limited efficacy of antibiotics, and the increasing incidence of multi‐drug resistant strains, novel therapeutics are urgently needed. An interesting new approach is to combine antibiotics with treatments that boost the natural ability of host cells to kill the bacteria, so‐called host‐directed therapeutics (HDT). Until now, this approach has not been explored in the context of nanoparticle drug delivery, which may offer several advantages. Here, the HDT drug everolimus and the antibiotic ((S)‐2‐nitro‐6‐((5‐((4‐(trifluoromethoxy)phenyl)ethynyl)pyridin‐2‐yl)methoxy)‐6,7‐dihydro‐5H‐imidazo[2,1‐b][1,3]oxazine, drug D) are focused on. Both of these agents, drug D and everolimus, are encapsulated into one single micelle (PeptoMicelles), resulting in formulation C, and compared with micelles containing only drug D (formulation A) or everolimus (formulation B). They are tested in Mtb‐infected primary macrophages and THP‐1 cells, and in the zebrafish model for TB (Mm infection) using embryo survival analysis. The results show that formulation C, containing drug D and everolimus in one micelle, is significantly more efficient in protecting cells and zebrafish embryos against infection than all other formulations, indicating an improved therapeutic efficacy of HDTs and antibiotics in a single polymeric micelle.

Investigating bacteria-induced inflammatory responses using novel endometrial epithelial gland organoid models

IntroductionThe endometrium plays a crucial role in early human pregnancy, particularly in embryo implantation, survival, and growth. However, invasion and infection by pathogens can lead to endometritis, infertility, and poor reproductive outcomes. Understanding the mechanisms of endometritis and its impact on fertility remains limited. An infection model using patient-derived endometrial epithelial gland organoids (EEGOs) was established to advance in vitro studies on endometritis and related infertility.MethodsAn EEGOs infection model was constructed and characterized from human endometrium, treating the organoids with estrogen and progesterone to observe changes in the proliferative and secretory phases. The organoids were infected with E. coli, and the release of inflammatory cytokines in the supernatant was detected using ELISA. RNA-seq was employed to analyze the differences before and after E. coli treatment, and differential gene mRNA expression was validated using real-time quantitative PCR. Additionally, the effect of E2 in alleviating inflammation was assessed through markers of receptivity (PAEP, LIF, ITGβ), proliferation (Ki67), and barrier repair (ZO-1).ResultsThe constructed human EEGOs exhibited long-term expansion capability, genetic stability, and characteristic hormonal responses, strongly expressing epithelial markers (MUC1, E-Cadherin). After E. coli infection, the expression levels of inflammatory cytokines TNF-α, IL-8, and IFN-γ increased significantly (P < 0.05). RNA-seq indicated that the MAPK signaling pathway was activated post-infection, with increased expression levels of heat shock proteins and transcription factor mRNA. E2 treatment post-infection significantly decreased the mRNA expression of inflammatory genes IL-1β, IL8, IL6 and TNF-α compared to the E. coli infected group (P < 0.05). Additionally, the expression of genes related to receptivity, proliferation, and barrier repair was enhanced in the E2-treated organoids.ConclusionsOur findings demonstrate that patient-derived EEGOs are responsive to bacterial infection and are effective models for studying host-pathogen interactions in bacterial infections. These organoids revealed the anti-inflammatory potential of E2 in alleviating E. coli-induced inflammation, providing insights into the mechanisms of endometritis and its impact on infertility. The study supports the use of EEGOs as valuable tools for understanding endometrial health and developing targeted treatments.

Influence of hormonal treatments on progesterone levels to enhance embryo survival and kidding rates in goats.

This study investigated the efficiency of different estrous synchronization programs and hormonal treatments in improving reproductive outcomes in goats. Conducted on a commercial farm in southern Thailand, the study used multiparous Shami and Anglo-Nubian breed goats. In experiment 1, goats were randomly allocated to two estrous synchronization treatments: 11-day (CI11D) and 13-day (CI13D) intravaginal progesterone implants, followed by artificial insemination (AI) with frozen-thawed semen. Various hormonal treatments (human chorionic gonadotropin (hCG), GnRH analogue, progestogen) were administered on day 5 post-AI to elevate progesterone levels. Experiment 2 assessed embryo survival after transferring early- and late-stage embryos, using GnRH analogue to increase progesterone levels in the recipient goats. Results showed that GnRH analogue significantly increased pregnancy rates, kidding rates, and the number of kids born in the CI13D group. Progesterone levels were higher in treated groups, particularly with GnRH analogue, though estradiol levels varied between synchronization protocols. Late-stage embryo transfers significantly improved pregnancy rates and reduced embryonic loss compared to early-stage transfers. GnRH analogue supplementation reduced early embryonic mortality, enhancing embryo survival and overall reproductive performance. This study demonstrates the efficacy of tailored estrous synchronization and hormonal treatments in optimizing goat reproductive outcomes, with significant implications for improving fertility management in commercial goat farming. Although no significant differences were observed in progesterone levels, the administration of GnRH analogue reduced early embryonic mortality and improved reproductive outcomes, demonstrating its potential to enhance embryo survival and reproductive performance in goats.

Comparison Between Electroporation at Different Voltage Levels and Microinjection to Generate Porcine Embryos with Multiple Xenoantigen Knock-Outs

In the context of xenotransplantation, the production of genetically modified pigs is essential. For several years, knock-out pigs were generated through somatic cell nuclear transfer employing donor cells with the desired genetic modifications, which resulted in a lengthy and cumbersome procedure. The CRISPR/Cas9 system enables direct targeting of specific genes in zygotes directly through microinjection or electroporation. However, these techniques require improvement to minimize mosaicism and low mutation rates without compromising embryo survival. This study aimed to determine the gene editing potential of these two techniques to deliver multiplexed ribonucleotide proteins (RNPs) to generate triple-knock-out porcine embryos with a multi-transgenic background. We designed RNP complexes targeting the major porcine xenoantigens GGTA1, CMAH, and B4GALNT2. We then compared the development of mosaicism and gene editing efficiencies between electroporation and microinjection. Our results indicated a significant effect of voltage increase on molecule intake in electroporated embryos, without it notably affecting the blastocyst formation rate. Our gene editing analysis revealed differences among delivery approaches and gene loci. Notably, employing electroporation at 35 V yielded the highest frequency of biallelic disruptions. However, mosaicism was the predominant genetic variant in all RNP delivery methods, underscoring the need for further research to optimize multiplex genome editing in porcine zygotes.

Effects of acute exposure to freshwater acidification on developing Oryzias latipes

Elevated CO2 in aquatic environments causes weak acidification. When exposed to weak acidification, regardless of life stage, most fishes undergo some degree of hypercapnia (elevation of CO2 in the bloodstream). Hypercapnia negatively affects physiological processes and embryo and larval fish are particularly vulnerable to rises in CO2. The aim of our study was to understand if weak acidification induced by elevated CO2 alters the physiology and behaviour of freshwater Japanese medaka (Oryzias latipes) embryos and larvae. To test this, we treated Japanese medaka embryos and larvae for 24 h with varying levels of weak acidification (pH 7.1 [~482 μatm], 6.4 [~2122 μatm], 6.1 [~3280 μatm], 5.8 [~5306 μatm], and 5.7 [~10,130 μatm]) at two developmental stages (72 hpf and 9 dpf). Following the treatment, heart rate, burst activity (movement within the egg), and survival of embryos were quantified. Swimming activity of hatched larvae was also measured. We observed a statistically significant 2-fold decline in heart rate of embryonic Japanese medaka as pH decreased (P < 0.01). We also found that survival of embryos significantly declined as acidification increased (P < 0.01). Behaviour of larval fish was significantly altered (P < 0.001) but not in a pH dependent manner. Our study suggests that weak acidification can cause negative effects to early life stage physiology and that behaviour can be altered. Our results suggest that if fish develop in weakly acidified freshwater there may be unfavourable impacts, including mortality at the most extreme levels.

Effects of intrauterine extracellular vesicle microRNAs on embryonic gene expression in low-fertility cows.

Embryo survival and pre-implantation development depend on uterine luminal fluid, which is believed to play a role in early embryonic death and infertility in cows. Extracellular vesicles (EVs) in the uterine luminal fluid contain microRNAs (miRNAs), crucial mediators of intercellular communication. miRNAs regulate conceptus-maternal interactions and participate in embryonic development by suppressing gene expression. Therefore, we hypothesized that miRNAs in the intrauterine EVs of low-fertility cows would hinder embryonic survival and development. EVs were collected from the bovine uterine luminal fluid of both normal- and low-fertility cows 7 days post-estrus. Small RNA-sequencing analysis of miRNAs isolated from these EVs identified eight miRNAs that were highly expressed in normal-fertility cows (normal-fertility miRNAs) and eight with elevated expression in low-fertility cows (low-fertility miRNAs). These two sets of miRNAs were transfected into hatched blastocysts via lipofection. RNA-seq following lipofection with low-fertility miRNAs identified 424 differentially expressed genes (DEGs) relative to the control; in contrast, following lipofection with normal-fertility miRNAs, seven DEGs were identified. Pathway analysis of the DEGs identified following lipofection with low-fertility miRNAs revealed substantial enrichment of mitogen-activated protein kinase (MAPK) signaling. Expression of activator protein 1 (AP1) and interferon-tau (IFNT) mRNA was significantly lower in the low-fertility miRNA transfection group than in the control. IFNT is essential for maternal pregnancy recognition. Therefore, miRNAs in intrauterine EVs from low-fertility cows at 7 days post-estrus may inhibit embryo development and suppress IFNT expression by altering MAPK signaling.

Trehalose polyphleates participate in Mycobacterium abscessus fitness and pathogenesis.

Mycobacteria produce a large repertoire of surface-exposed lipids with major biological functions. Among these lipids, trehalose polyphleates (TPPs) are instrumental in the infection of Mycobacterium abscessus by the therapeutic phage BPs. However, while the biosynthesis and transport of TPPs across the membrane by MmpL10 have been reported, the role of TPPs in host infection remains enigmatic. Here, we addressed whether the loss of TPPs influences interactions with macrophages and the virulence of M. abscessus. As anticipated, the deletion of mmpL10 in smooth (S) and rough (R) variants of M. abscessus abrogated TPP production, which was rescued upon gene complementation. Importantly, infection of human THP-1 cells with the mmpL10 mutants was associated with decreased intramacrophage survival and a reduced proportion of infected cells. The rough mmpL10 mutant showed an impaired capacity to block phagosomal acidification and was unable to co-localize with Galectin-3, a marker of phagosomal membrane damage. This suggests that TPPs participate, directly or indirectly, in phagolysosomal fusion and in phagosomal membrane damage to establish cytosolic communication. The TPP defect that affects the fitness and virulence of M. abscessus was further demonstrated in zebrafish embryos using a rough clinical strain resistant to phage BPs and harboring a frameshift mutation in mmpL10. Infection with this strain was correlated with a slight decrease in embryo survival and a reduced bacterial burden as compared to the corresponding parental and complemented derivatives. Together, these results indicate that TPPs are important surface lipids contributing to the pathogenicity of M. abscessus.IMPORTANCETrehalose polyphleates (TPPs) are complex lipids associated with the mycobacterial cell surface and were identified 50 years ago. While the TPP biosynthetic pathway has been described recently, the role of these lipids in the biology of mycobacteria remains yet to be established. The wide distribution of TPPs across mycobacterial species suggests that they may exhibit important functions in these actinobacteria. Here, we demonstrate that Mycobacterium abscessus, an emerging multidrug-resistant pathogen that causes severe lung diseases in cystic fibrosis patients, requires TPPs for survival in macrophages and virulence in a zebrafish model of infection. These findings support the importance of this underexplored family of lipids in mycobacterial pathogenesis.

Сравнително проучване на морфологичните качества и люпимостта на яйцата от чисти линии и кръстоски от общоползвателно и яйценосно направление

During the recent decades, the scientific community turned its attention towards the dual purpose lines and crosses that might offer a suitable solution for two problems in poultry breeding - the use of male chickens for meat and the use of female chickens for rearing in alternative types of systems for egg production. Hence, a study with four groups of birds: a dual purpose cross „Salmon“, a dual purpose purebred line „BB“, a laying line „L“ and a laying hybrid line „ISA-Brown“ was conducted. The aim was to compare the morphological traits, as well as the fertility and the hatchability of the eggs, obtained from „ISA-Brown“, compared to the dual purpose „Salmon“ reared at IAS-Kostinbrod, and the local dual purpose and laying lines involved in its creation. As a result of the conducted research, we can conclude that the selection for high performance, larger eggs and the presence of heterosis in hybrids has to some extent a positive influence on most of the morphological traits determining the quality of the eggs produced. In addition, negative consequences for their hatchability, embryo survival and some morphological parameters such as increased width of the egg and increased diameter of the yolk were also observed. On the other hand, in the stock hybrids, compared to the purebred lines and the dual purpose crosses, this can also be considered as a sign of deterioration of the nutritional quality of the eggs.

Between Life and Death: Sea Urchin Embryos Undergo Peculiar DNA Fragmentation after Exposure to Vanadium, Cadmium, Gadolinium, and Selenium.

Exogenous DNA damage represents one of the most harmful outcomes produced by environmental, physical, or chemical agents. Here, a comparative analysis of DNA fragmentation was carried out on Paracentrotus lividus sea urchin embryos exposed to four common pollutants of the marine environment: vanadium, cadmium, gadolinium and selenium. Using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, fragmented DNA was quantified and localized in apoptotic cells mapping whole-mount embryos. This is the first study reporting how different chemicals are able to activate distinctive apoptotic features in sea urchin embryos, categorized as follows: (i) cell-selective apoptosis, showing DNA fragmentation restricted to a subset of extremely damaged cells, acting as an embryo survival mechanism; or (ii) total apoptosis, with fragmented DNA widespread throughout the cells of the entire embryo, leading to its death. Also, this is the first report of the effects of Se exposure on P. lividus sea urchin embryos. These data confirm the TUNEL assay as the most suitable test to study DNA fragmentation in the sea urchin embryo model system. Taken together, this research highlights embryos' ability to find alternative pathways and set physiological limits for development under stress conditions.

Characterisation of Fatty Acid Profiles in Tasmanian Atlantic Salmon Muscle, Oocytes and Eggs in Differing Fully Commercial Settings

ABSTRACTTasmanian Atlantic salmon broodstock may be conditioned in flow‐through (FT) systems where water quality fluctuates and temperatures approach their upper limit of thermal tolerance, or in recirculating aquaculture systems (RAS) where conditions are comparatively cool and stable. The impact of broad conditioning approach on the molecular cargo of salmon eggs has never been explored in dynamic commercial settings. Therefore, this study aimed to characterise the reproductive features of broodstock conditioned using different commercial approaches and determine whether the fatty acid (FA) profiles of muscle, oocytes and eggs varied between groups during vitellogenesis and at stripping. The collective conditioning approach had a marked impact on the somatic and reproductive dynamics of female salmon broodstock, and the eggs produced by each group of fish were fundamentally different in terms of their FA composition. Of particular note is the increased maternal investment of saturated FAs (mg.g−1) and monounsaturated FAs (percent and mg.g−1) by FT‐based eggs, and the higher n3 polyunsaturated FA (percent and mg.g−1), docosahexaenoic acid (DHA, percent and mg.g−1), arachidonic acid (ARA, percent), eicosapentaenoic acid (EPA, percent), n3:n6 ratio and ARA:EPA ratio of RAS‐based eggs. These metrics combined with the apparently higher prevalence of neural development and survival of RAS embryos suggest that RAS eggs were of higher quality. On the other hand, there is evidence to suggest that FT broodstock attempt to prepare their offspring for a (presumably) challenging early life environment by producing larger eggs that are proportionally rich in short‐chain FAs and contain a higher total quantity of FAs per egg.

Advancing Maternal Transfer of Organic Pollutants across Reptiles for Conservation and Risk Assessment Purposes.

Embryonic exposure through maternally transferred pollutants can affect embryo vitality, survival, and health. Reptiles face global declines and are sensitive to embryonic pollutant exposure. Yet, they are often neglected in pollution risk assessment and conservation. We analyzed maternal transfer of organic pollutants in reptiles through a systematic extraction, homogenization, and integration of published data on organic pollutants measured in mother-egg pairs into a comprehensive database (DOI:10.5281/zenodo.10900226), complemented with molecular physical-chemical properties of the pollutants. Over four decades, 17 publications provided 19,955 data points shifting from legacy to emerging contaminants although research on newer contaminants lags regulatory and societal demands. Challenges including taxonomic bias, heterogeneity in sampled tissues, and 73% of censored data complicate comparative analyses. However, significant opportunities were identified including the use of the turtle Malachlemys terrapin and snake Enhydris chinensis as flagship species where a large amount of data is available across tissues (allowing investigation into physiological relations) and compounds (allowing insights into maternal transfer across the chemical universe). Data on other freshwater and marine turtles provide the possibility of exploring taxonomic patterns in this subgroup. The analysis, integrated database, and discussion present opportunities for research in an era where science needs to achieve more with limited wildlife data.

Interacting phenotypic plasticities: Do male and female responses to the sociosexual environment interact to determine fitness?

Socially induced plasticity in reproductive effort is a widely documented phenomenon. However, few empirical studies have examined how male and female plastic responses to the social environment might interact in determining fitness outcomes. In field crickets, Teleogryllus oceanicus, males respond to rival song by increasing expenditure on seminal fluid proteins that enhance competitive fertilization success at the cost of reduced embryo survival. It remains unknown whether plastic responses in females could moderate the effects of male competitiveness on offspring performance. Here we used a fully factorial design to explore the interacting effects on fitness of male and female plasticity to the sociosexual environment. We found that female crickets exposed to male song increased the number of eggs produced during early life reproduction, which came at a cost of reduced offspring size. There was evidence, albeit weak, that interacting effects of male and female sociosexual environment contributed to variation in the hatching success of eggs laid by females. Lifetime offspring production was unaffected by the sociosexual environments to which upstream male and female plastic responses were made. Our data offer a rare test of the theoretical expectation that male and female plasticities should interact in their effects on female fitness.

PSII-14 Climate and conception: Investigating the resilience of equine embryo survival to ambient temperatures in southeast Texas

Abstract Amidst the unique challenges posed by the humid and hot climate of southeast Texas, this study aimed to assess the establishment of pregnancy from transferred equine embryos, hypothesizing that an increase in ambient temperature would lead to a decrease in pregnancy establishment. To achieve this, we utilized 17 embryo donors and 46 recipient mares (3 to 9 yr of age), all anatomically and clinically normal with no history of reproductive problems. Breeding records for 49 transferred d 8, grade-1 embryos from a single private horse farm in southeast Texas, along with the reported high and low ambient temperatures on the day of transfer, were analyzed using logistic regression via PROC GLIMMIX in SAS v9.4 with fixed effects for the reported high and low ambient temperatures on the day of transfer, with a random statement for donor and sire. The pregnancy rate for all embryo transfers was 79%. Surprisingly, our results revealed that neither the reported high ambient temperatures (P = 0.16) nor reported low ambient temperatures (P = 0.19) significantly influenced the likelihood of pregnancy establishment observed at d 14 post-ovulation of transferred equine embryos. Furthermore, no statistically significant influence (P &amp;gt; 0.05) on pregnancy establishment was observed when embryo transfers were conducted outside of the thermal neutral zone (5°C to 20°C). Within the context of our study, these results suggest that environmental temperatures in southeast Texas do not significantly impact equine pregnancy outcomes.

Co-infection of Liburna oophaga sp. nov. and Ikanecator primus on cuttlefish (Sepia pharaonis) eggs and the effectiveness of peracetic acid as a treatment

The cuttlefish Sepia pharaonis species complex is emerging as a promising set of organisms for research in neuroscience, the behavioral sciences, and commercial aquaculture. At the same time, information about pathogens and diseases that could affect cuttlefish cultivation in intensive aquaculture settings remains limited. Our study has identified two species of parasite, the protozoan Liburna oophaga sp. nov. and the metazoan Ikanecator primus, that co-infect cuttlefish eggs, increasing mortality and reducing hatching rates. L. oophaga sp. nov. is reported here for the first time to enhance mortality during the incubation period by inducing deformity in cuttlefish eggs. We investigated the application of peracetic acid to parasite elimination during cuttlefish egg incubation. When cuttlefish eggs were treated with a peracetic acid containing product (PAA-product); 35 mg/L PAA + 15 mg/L H2O2, L. oophaga on the surfaces of the eggs were eliminated within 10 min. PAA-product; 70 mg/L PAA + 30 mg/L H2O2 was required to achieve the same effect for I. primus. Immersion treatment with PAA-product at 70 mg/L PAA + 30 mg/L H2O2 reduced parasitic load and improved survival of cuttlefish embryos and hatchling size, demonstrating that PAA product can inhibit and control parasitic co-infections in cephalopod culture.

Effects of parity and early pregnancy on peripheral blood leukocytes in dairy cattle

Subfertility remains a major problem in the dairy industry. Only 35-40% of high-yielding dairy cows and 55-65% of nonlactating heifers become pregnant after their first service. The immune system plays a critical role in the establishment of pregnancy. However, it can also create challenges for embryo survival and contribute to reduced fertility. We conducted 2 separate experiments to characterize changes in subsets of peripheral blood leukocytes (PBL) and their phenotype over the estrous cycle and early pregnancy in heifers and cows. We used flow cytometry and RT-qPCR to assess protein and mRNA expression of molecules important for immune function. We observed that CD14+ monocytes and CD3+ T cells tended to be affected by pregnancy status in heifers, whereas CD8B+ lymphocytes and NCR1+ natural killer (NK) cells were affected during early pregnancy in cows. Changes in expression of immune function proteins appeared to be greater in heifers than cows. To compare the most striking differences between heifers and cows observed in the initial experiments, we conducted a third experiment where PBL sampled from heifers and cows were simultaneously collected and analyzed under the same experimental conditions. Our results indicate that, compared with heifers, cows had greater mRNA expression of proinflammatory cytokines (IFNG and IL6) and AHR protein along with greater percentage of MM20A+ neutrophils and myeloid cells expressing SIRPA, ITGAM and ITGAX. Moreover, animals that failed to become pregnant showed altered expression of anti-inflammatory molecules compared with cyclic and pregnant animals. Overall, these findings support the hypothesis that early pregnancy signaling alters the phenotype of immune cells in the peripheral blood and that there are differences in the peripheral immune response to pregnancy between cows and heifers. Because cows have lower conception rates than heifers, it is possible that a more proinflammatory immune status in peripheral blood may play a role in embryo loss.

Evaluating the biomedical and environmental safety of selenium nanoparticles synthesized from black pepper seed extract

In recent times, sustainable synthesis of selenium nanoparticles (Se NPs) has seen a significant rise in research, focusing primarily on microbial and plant polysaccharide-based approaches for various bioapplications. However, there are limited studies on the use of extracts from herbal spice plants as capping agents for Se NPs. Therefore, this study investigates the pioneering synthesis of Se NPs using black pepper (Piper nigrum) seed extract (PSe NPs) and assesses their antioxidant properties, hemolytic potential, antibacterial and antibiofilm activities, cytotoxicity, phytotoxicity, in vivo toxicity in zebrafish embryos, and their effectiveness in prolonging the shelf life of food materials. The green-synthesized PSe NPs were thoroughly physicochemically characterized and found to be sphere-shaped with a medium diameter of 10–100 nm. The PSe NPs exhibited substantial antibacterial effects, being more effective against S. aureus (Gram-positive)than against S. typhi (Gram-negative). Furthermore, PSe NPs significantly inhibited MRSA biofilm growth at 500 μg/mL and showed notable antioxidant activity compared to standard gallic acid. Also at 500 μg/mL, PSe NPs induced significant hemolysis, whereas inferior concentrations did not. Cytotoxicity evaluations using the MTT assay revealed that PSe NPs significantly reduced the viability of human normal embryonic kidney (HEK 293) cells and human cervical cancer (HeLa) cells after 24 hours of treatment at concentrations of 0.1–500 μg/mL. This cytotoxic effect was associated with apoptosis, as shown by acridine orange/ethidium bromide staining. In phytotoxicity assays, PSe NPs revealed significant effects on seed germination and root and shoot growth of the mung bean (Vigna radiata). The zebrafish embryotoxicity assay indicated that PSe NPs at 500 µg/mL affected embryo morphology, mortality and survival rates 96 hours after fertilization. Furthermore, the application of PSe NPs gel coatings effectively reduced weight loss and extended the shelf life of fruits such as apples and pears, maintaining their natural appearance.

Essential Roles of PIEZO1 in Mammalian Cardiovascular System: From Development to Diseases.

Mechanical force is the basis of cardiovascular development, homeostasis, and diseases. The perception and response of mechanical force by the cardiovascular system are crucial. However, the molecular mechanisms mediating mechanotransduction in the cardiovascular system are not yet understood. PIEZO1, a novel transmembrane mechanosensitive cation channel known for its regulation of touch sensation, has been found to be widely expressed in the mammalian cardiovascular system. In this review, we elucidate the role and mechanism of PIEZO1 as a mechanical sensor in cardiovascular development, homeostasis, and disease processes, including embryo survival, angiogenesis, cardiac development repair, vascular inflammation, lymphangiogenesis, blood pressure regulation, cardiac hypertrophy, cardiac fibrosis, ventricular remodeling, and heart failure. We further summarize chemical molecules targeting PIEZO1 for potential translational applications. Finally, we address the controversies surrounding emergent concepts and challenges in future applications.

The reproductive costs of bitterling fish and zebra mussel parasitism to a unionid mussel

Unionid mussels are one of the most vulnerable groups of animals, with multitude of factors responsible for the decline of their populations in the freshwater ecosystems. Many of those causes stem from interspecific relationships. Direct data on the reproductive costs incurred to unionids through parasitism by fish and other mussel species are incomplete. We used field experiments to test whether reproduction of the swollen river mussel (Unio tumidus) is affected by parasitism by the European bitterling fish (Rhodeus amarus) and zebra mussel (Dreissena polymorpha). We documented that bitterling brood can develop in the gills of female, male and hermaphrodite mussels. Our research showed that male mussels carried more fish offspring than female U. tumidus and bitterling embryo survival appeared higher in the outer gill demibranchs. Parasitised female U. tumidus suffered reduced fecundity in terms of a reduced number of mature ovulated oocytes and increased oocyte degeneration. Our data lend clear support to the hypothesis that bitterling fish embryos parasitizing mussel gills and epibiont zebra mussel fouling unionid shells represent a major cost to the reproductive success of their host mussels. Extended gonadal activity (multiple spawning) of U. tumidus spreads their spawning time over a longer period, which may mitigate the costs of parasitism.

Transgenerational effects of the levonorgestrel-based birth control pill in zebrafish offspring

The consumption of hormone-derived medicines, such as levonorgestrel (LNG), is increasing worldwide, and its discharge into the environment reaches non-target organisms. In our previous study, we exposed the parental generation of zebrafish to environmentally relevant concentrations of LNG during the developmental phase. Subsequently, they had grown in a tank with clean water until adulthood. Now, we allowed this parental generation to reproduce to obtain F1 progeny unexposed to LGN, in order to analyze the transgenerational effects of parental LNG exposure on the survival and hatching of unexposed F1 embryos and the stress and behavior of F1 larvae. Here, we found decreased survival rates with higher LNG concentrations, providing a transgenerational effect. This highlights the environmental impact of exposure to LNG, causing damage at the individual and population level and affecting the next generation at the beginning of development, impacting qualities in the survival of the species.

Essential roles of histone lysine methyltransferases EZH2 and EHMT1 in male embryo development of Phenacoccus solenopsis

Paternal genome elimination (PGE) is an intriguing but poorly understood reproductive strategy in which females are typically diploid, but males lose paternal genomes. Paternal genome heterochromatin (PGH) occurs in arthropods with germline PGE, such as the mealybug, coffee borer beetles, and booklice. Here, we present evidence that PGH initially occurs during early embryo development at around 15 h post-mating (hpm) in the cotton mealybug, Phenacoccus solenopsis Tinsley. Transcriptome analysis followed by qPCR validation indicated that six histone lysine methyltransferase (KMT) genes are predominantly expressed in adult females. We knocked down these five genes through dsRNA microinjection. We found that downregulation of two KMT genes, PsEZH2-X1 and PsEHMT1, resulted in a decrease of heterochromatin-related methylations, including H3K27me1, H3K27me3, and H3K9me3 in the ovaries, fewer PGH male embryos, and reduced male offspring. For further confirmation, we obtained two strains of transgenic tobacco highly expressing dsRNA targeting PsEZH2-X1 and PsEHMT1, respectively. Similarly, fewer PGH embryos and fewer male offspring were observed when feeding on these transgenic tobacco plants. Overall, we present evidence that PsEZH2-X1 and PsEHMT1 have essential roles in male embryo survival by regulating PGH formation in cotton mealybugs.