Reproduction最新文献

Preeclampsia is a life-threatening pregnancy complication, resulting in >70,000 maternal deaths and >500,000 fetal and neonatal deaths worldwide each year. Despite its prevalence and devastating consequences, there is still no medical therapy to halt disease progression and thus a vital need for novel treatments remains. The urgent demand for effective therapies for preeclampsia first necessitates a deeper understanding of the pathogenesis that underpins the condition. Understanding the coordinated interactions between the maternal vasculature and circulating pathogenic factors released from the placenta is a key focus in preeclampsia research. Extracellular vesicles are membrane-bound vesicles that act as essential mediators of cell-cell signalling. Extracellular vesicles are proposed to play a crucial role as regulators in placental-maternal vascular communication. This -review explores contemporary literature investigating the role of placental-derived extracellular vesicles and their contribution to preeclampsia. Importantly, we provide a summary of key literature that should assist decision making for selection of models to study the role of extracellular vesicles in preeclampsia progression and their potential as therapeutic targets.

This study investigates whether follicle-stimulating hormone (FSH) induces ferroptosis in TM4 Sertoli cells and mouse testes, and identifies potential mitigating factors. TM4 cells and mice were treated with FSH, and assessments included cell viability, testicular histology, and key ferroptosis markers (ferrous iron, malondialdehyde malondialdehyde, glutathione glutathione). Molecular expression was analyzed via quantitative real-time PCR and western blot. The role of ferroptosis was further examined using the inhibitor ferrostatin-1 (Fer-1). RNA sequencing was employed to explore underlying mechanisms, and functional validation was performed through knockdown and overexpression of the identified regulator, fibroblast growth factor 21 (FGF21). Our results demonstrate that FSH exposure induces ferroptosis in both TM4 Sertoli cells and mouse testes. This is evidenced by decreased protein levels of the ferroptosis suppressors SLC7A11 and FSP1, increased levels of the stress-response proteins FTH1 and HO1, elevated ferrous ion and malondialdehyde content, and reduced glutathione. This ferroptotic cell death may represent a key mechanism contributing to FSH-associated testicular damage. Notably, the ferroptosis inhibitor ferrostatin-1 effectively mitigated this process in TM4 cells. Transcriptomic analysis not only confirmed FSH-induced ferroptosis but also identified FGF21 as a potential modulator. Knockdown of FGF21 promoted ferroptosis, whereas supplementation with exogenous FGF21 alleviated FSH-induced ferroptosis, suggesting a novel inhibitory role for FGF21 in this pathway. In summary, our findings establish that FSH can induce testicular ferroptosis and identify FGF21 as a potential endogenous mitigator of this effect. This highlights FGF21 as a promising therapeutic target for preventing or treating FSH-induced testicular damage.

The epididymis is a highly specialized organ essential for promoting the post-testicular functional maturation of spermatozoa, a process underpinning male fertility. This review examines the latest proteomics advances that have been used to unravel the complex molecular landscape of the epididymis, revealing the dynamic protein networks that shape sperm function beyond their genomic and transcriptomic blueprints. Here, we highlight how high-resolution mass spectrometry has helped to map the proteomic signatures of epididymal tissue, luminal extracellular vesicles (epididymosomes), and spermatozoa at different maturation stages, pinpointing key regulators of motility, capacitation, fertilization competence, and immune regulation. However, critical knowledge gaps remain, including deep protein characterization of the cytoplasmic droplet, epididymal fluid, and relatively underexplored anatomical tissue segments such as the corpus and cauda epididymis. We discuss how integrating global proteomic insights with complementary omics, single-cell proteomics, and advanced imaging is poised to reveal the spatial and temporal refinement of the sperm proteome, providing insights into how its disruption may contribute to idiopathic infertility. To promote data accessibility and accelerate discovery in epididymal biology, we introduce ShinyEpididymis (https://reproproteomics.shinyapps.io/ShinyEpididymis/), an interactive, web-based resource integrating publicly available proteomic datasets from spermatozoa, epididymosomes, and epididymal tissue. This platform enables researchers to rapidly query proteins of interest, explore spatial patterns of expression, and identify potential biomarkers or therapeutic targets. By consolidating current knowledge and defining future priorities, this review positions proteomics at the forefront of understanding epididymal biology, emphasising its clinical relevance and untapped potential for diagnosing and treating male infertility.

Preimplantation embryogenesis requires precise synchronization of transcriptional activation, mRNA export and translation, and metabolic reprogramming to sustain developmental requirements. Nuclear cap-binding protein 1 (NCBP1), a conserved subunit of the cap-binding complex, has established roles in mRNA processing and export in somatic cells, but its potential functions in preimplantation embryogenesis remain undefined. The spatiotemporal expression dynamics of Ncbp1 were explored on multiple levels. After microinjecting interfering RNA at zygotic stage to knockdown Ncbp1, embryonic developmental competence was evaluated. Co-injection of small interfering RNA and in vitro transcribed Ncbp1 mRNA into the zygote was used to rescue the knockdown phenotype. Further, poly-adenylated RNA-fluorescence in situ hybridization, RNA sequencing, and quantitative proteomics were used to investigate the effects of Ncbp1 knockdown. In addition, oleic acid (OA) supplementation was used to rescue developmental abnormalities. NCBP1 exhibited dynamic spatiotemporal expression coinciding with nuclear-to-cytoplasmic translocation of protein from morula stage. Depletion of Ncbp1 caused morula arrest or fragmentation, accompanied by nuclear poly-adenylated RNA retention and down-regulation of lipid metabolic pathways, notably, stearoyl-CoA desaturase 1 (SCD1), a key enzyme generating monounsaturated OA. Exogenous OA supplementation partially rescued blastocyst formation, implicating NCBP1 in the regulation of SCD1-OA-mediated metabolic homeostasis during morula-to-blastocyst transition. This study illustrates NCBP1 as a mediator that regulates RNA export and lipid homeostasis during early mouse embryo development. Especially NCBP1 regulates the SCD1-OA metabolic pathways, ensuring metabolic flexibility essential for successful morula-to-blastocyst transition, thereby providing new insights into the molecular basis of embryonic developmental competence.

Uterine smooth muscle undergoes electrical, chemical, and mechanical transients during phasic contraction. The spread of electrical activity precedes the mechanical contraction of muscle fibers, and the initiation and coordination of these events is critical to uterine function, in both pregnancy and non-pregnancy. Characterization of non-pregnant activity is scarce, and understanding of the relationship between electrical and mechanical events is not well understood. Isolated uterine horns from virgin female Wistar rats were used in this study. Electrical and mechanical activity were assessed ex vivo using high-resolution surface electrodes and motion tracking. Electrophysiology data were analyzed to provide frequency and timing metrics; displacement and velocity were quantified from the motion-tracking data. Major coordinated electrical events were correlated with longitudinal displacement of tissue markers, with the peak displacement occurring at or near the point of electrical signal initiation. Overall, 82% of electrical slow wave events were associated with mechanical displacement of tracking points. Minor uncoordinated electrical events were accompanied by little or no displacement, indicating that a level or coordination is required to achieve tissue-level contraction. Electrical slow wave propagation occurred at a higher speed (0.60 ± 0.04 mm/s) than mechanical tissue displacement velocity (0.42 ± 0.39 mm/s, p = 0.0024), and electrical propagation speed was lowest during metestrus when compared with estrus (p = 0.0042) and diestrus (p = 0.0066). This technique advances our understanding of the electro-mechanical properties of the non-pregnant rat uterus and provides an avenue to measure electrical-mechanical coupling in ex vivo uterine tissue preparations.

The pronuclei in human one-cell stage embryos (zygotes) contain several nucleoli called nucleolus precursor bodies (NPBs). Based on their number and distribution, it is possible to predict the developmental potential of the zygote. Recently, it has been demonstrated that the speed of NPBs movement in pronuclei may also indicate how the embryo will develop, as well as its chromosomal constitution (euploidy versus aneuploidy). These observations, however, do not elucidate the mechanisms behind these processes, and a deeper understanding will certainly be important for the more efficient production of healthy human embryos.

Trehalose is a disaccharide known for protecting cells against desiccation and freezing. However, mammalian cells lack endogenous trehalose transporters, limiting its intracellular utility. In this study, we examined whether transient expression of pvTret1, a trehalose transporter from the desiccation-tolerant midge Polypedilum vanderplanki, could enable trehalose uptake in mouse embryos without impairing development. We synthesized mRNA encoding pvTret1-eGFP and microinjected it into mouse zygotes. Fluorescence microscopy confirmed plasma membrane localization, and AlphaFold modeling indicated that eGFP fusion did not alter protein structure. Embryos expressing pvTret1 developed to the blastocyst stage and produced live offspring at rates comparable to sham-injected controls, suggesting that transient expression does not compromise -developmental potential. LC-MS analysis revealed that pvTret1-expressing embryos took up trehalose in a concentra-tion--dependent manner following 20-min exposure to trehalose-containing medium, and effectively cleared intracellular -trehalose within 20 min of removal, indicating functional, bidirectional transport. In contrast, control embryos exhibited shrinkage under hyperosmotic conditions. Embryos with intracellular trehalose also showed improved post-warming survival after vitrification in dimethyl sulfoxide-free, trehalose-based cryoprotectant solutions. Although optimization is ongoing, these findings highlight the potential utility of pvTret1-mediated trehalose uptake in developing low-toxicity embryo preservation methods. This is the first demonstration that pvTret1 can function in mammalian embryos, enabling transient intracellular trehalose accumulation without transgenesis. Future work will focus on refining trehalose loading protocols and advancing vitrification or dry-preservation strategies for reproductive and biomedical applications.

Humans and rhesus macaques are known to express two molecular forms of gonadotropin-releasing hormone (GnRH-I and GnRH-II), which appear to differentially contribute to the regulation of the menstrual cycle. Specifically, there is evidence to suggest that GnRH-I is the primary mediator of negative estrogen feedback to the hypothalamus and pituitary gland, while GnRH-II is the primary mediator of the positive feedback that stimulates the preovulatory surge of luteinizing hormone. Therefore, it is plausible that selective silencing of GnRH-II would block ovulation and lay the platform for development of a novel contraceptive. To test this possibility female rhesus macaques were actively immunized against GnRH-II (and/or GnRH-I), and serum estradiol and progesterone concentrations were monitored for an additional ∼2.5 years. Despite multiple booster immunizations every ∼6 weeks, and elevated GnRH antibody titers, none of the animals ceased ovulating (i.e., revealed by monthly peaks of serum progesterone concentrations followed by menstruation). Taken together, these findings question the efficacy of GnRH vaccines as a stratagem for selectively blocking ovulation in humans. However, they do not negate the potential value of pharmacological interventions aimed at selectively silencing GnRH-II function and its involvement in stimulating the preovulatory luteinizing hormone surge.

Diisononyl phthalate (DiNP), a plasticizer increasingly replacing di(2-ethylhexyl) phthalate, is an endocrine-disrupting chemical linked to female reproductive harm. Ingestion is the most common route of DiNP exposure, making the gastrointestinal tract and gut microbiome a direct target for endocrine-disrupting chemical exposure. This study examined the effects of acute DiNP exposure either in the absence or presence of a gut microbiome on uterine development. Female C57Bl/6 germ-free (-microbiome) 40-day-old mice were orally dosed, over 3 days, with either sterile phosphate-buffered (n = 8) to remain germ-free (GF, -microbiome) or with colon contents (n = 10) to develop a gut-microbiome (+microbiome). This was followed by a 10-day period where half of the -microbiome and +microbiome mice were orally dosed with corn oil while half were orally dosed with 200 μg/kg/day DiNP. The control group were specific pathogen-free conventionally housed mice born with a microbiome. Mice were euthanized in diestrus at the end of the 10 days. Uteri were collected for histological analyses. Uterine development was significantly delayed in GF mice, regardless of later microbiome reintroduction or DiNP exposure. Key findings included reduced uterine diameter, stroma area, and gland number, and thinner myometrial layers. Endometrial stromal cell proliferation was also lower in GF mice. DiNP exposure alone showed no significant effects. Estradiol levels and ovarian follicle counts were similar across groups, but GF mice had fewer, smaller litters in fertility tests. The study highlights that the gut microbiome critically influences postnatal uterine development, with its absence leading to persistent structural deficits. DiNP, at the tested dose, did not exacerbate these effects.

Glucose transport across the placenta is essential for fetal growth and development. Glucose transporter 1, encoded by the SLC2A1 gene, plays a central role in mediating maternal-fetal glucose exchange. Dysregulation of placental glucose transport is implicated in pregnancy-related complications, such as preeclampsia and fetal growth restriction; however, the mechanistic role of SLC2A1 in trophoblast function remains poorly defined. To functionally validate the role of SLC2A1 in human trophoblasts, we used clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9-mediated knockout of the SLC2A1 gene, enabling complete and permanent loss of SLC2A1 expression. In the resulting SLC2A1 knockout human trophoblast HTR8/SVneo cells, SLC2A1 depletion induced a metabolic shift from glycolysis to oxidative phosphorylation, leading to increased mitochondrial respiration, ATP production, mitochondrial calcium overload, and elevated mitochondrial reactive oxygen species generation. These changes were accompanied by enhanced endoplasmic reticulum stress, as shown by the upregulation of p-PERK, IRE1α, and GRP78, as well as increased autophagic activity indicated by LC3B-II and p62 accumulation. Notably, mTOR signaling was also upregulated, suggesting a feedback loop that regulates autophagy. The loss of SLC2A1 impaired the PI3K/AKT pathway, reduced trophoblast migration and 3D spheroid formation, and disrupted epithelial-mesenchymal transition-like properties. These findings demonstrate that SLC2A1 is essential for maintaining trophoblast energy homeostasis, redox balance, and invasive capacity; its deficiency triggers mitochondrial and endoplasmic reticulum stress responses that may contribute to placental dysfunction during early pregnancy.