Biology of Reproduction最新文献

In mammals, seminal plasma is a complex fluid surrounding spermatozoa, composed of secretions from the male reproductive tract. It plays a crucial role in modulating sperm function, but it remains unclear whether the components that regulate sperm physiology, travel freely or within extracellular vesicles secreted by the reproductive tract and accessory glands. This study evaluated three methodologies-ultracentrifugation (UC), size-exclusion chromatography (SEC), and polyethylene glycol precipitation (PEG)-for isolation of ram seminal plasma extracellular vesicles enriched fractions (SP-EVs), assessing their efficiency in terms yield, morphology, protein profile, and functionality. Western blot confirmed the presence of EV-specific markers (CD9, CD63, and HSP70), minimal cytoplasmic and lipoprotein contamination. SEC, particularly the second fraction (P2), yielded SP-EVs with conserved morphology, apparently reduced aggregation, and a unique protein profile enriched in low molecular weight proteins, compatible with most capacitation-modulating proteins. In contrast, UC and PEG resulted in higher particle concentration and aggregation. By CFSE labeling of SP-EVs, all preparations exhibited a targeted binding pattern to spermatozoa, with distinct patterns localized to midpiece, head and post-acrosomal regions. Additionally, western blot analysis showed that SP-EVs transport and transfer binder of sperm proteins (RSVP20 and RSVP14) to spermatozoa, with RSVP20 showing the highest incorporation, particularly from the P2 fraction. SPINK3, despite being detected in SP-EVs, was not incorporated, indicating selective protein delivery. These findings may be important to understand the role of seminal plasma extracellular vesicles on sperm, and significant for improving the efficiency of reproductive biotechnologies, as these ram SP-EVs enrichment fractions can deliver functional proteins to spermatozoa.

European research has played a pioneering role in advancing reproductive biology and embryology, contributing transformative innovations in assisted reproductive technologies (ARTs). Landmark achievements-from the development of artificial insemination and sperm cryopreservation to in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI)-have dramatically improved fertility outcomes in both humans and animals. These breakthroughs have enabled increased reproductive success, genetic gain, and productivity in livestock, while also addressing the global burden of infertility in humans. Central to recent progress is the application of high-throughput single-cell RNA sequencing, which has revealed conserved and species-specific molecular pathways during early embryonic development. Parallel efforts have led to the development of synthetic embryo models, showcasing the self-organizing capacity of pluripotent cells and offering alternatives to natural embryo studies, particularly in species where embryo availability is limited. These models recapitulate key developmental milestones and are informed by high-resolution single-cell atlases of multiple species, many of which were created in European institutions. While notable advances have been made, challenges persist in achieving reproducibility and more robust implantation models. Nevertheless, the integration of genomic, stem cell, and bioengineering technologies continues to expand the frontiers of reproductive biology. This review highlights Europe's significant contributions to embryological research and outlines the emerging tools and challenges shaping the future of reproductive science.

Anti-Müllerian hormone is a factor secreted by granulosa cells of growing follicles that regulates many aspects of ovarian function, including primordial follicle activation and early follicle development through inhibitory feedback. Treatment with exogenous anti-Müllerian hormone at supraphysiological levels can inhibit follicular development and prevent ovulation in mice, rats, and cats. Single-cell transcriptomic analysis of ovaries from mice treated postnatally with anti-Müllerian hormone identified a distinct quiescence signature in granulosa cells. Herein, we further characterized the anti-Müllerian hormone -induced quiescent cell state and drew parallels to developmental senescence by describing a reversible "quiescence-associated secretory phenotype." This state was characterized by the induction of several hallmarks of senescence, including inhibition of proliferation (reduced KI67), upregulation of markers of senescence (Cdkn1a, Fn1, Cebpb, Timp3), and chemokines and their receptors (Cxcl14, Cxcl12, Ccl12, Ccl21, Cxcr2). However, quiescence-associated secretory phenotype did not recapitulate other senescence hallmarks, such as activation of beta-galactosidase activity or permanent exit from the cell cycle. Similar to classical senescence, we found that anti-Müllerian hormone induced quiescence-associated secretory phenotype, in part, through the upregulation of cyclin-dependent kinase inhibitors (Cdkn1a, Cdkn1b, and Cdkn1c) in granulosa cells and oocytes of mouse and human ovaries. Finally, we showed that the quiescence-associated secretory phenotype state could be reversed within a month of discontinuing anti-Müllerian hormone treatment. This recovery was marked by the return of growing primary, secondary, and antral follicles. In conclusion, these data suggest that anti-Müllerian hormone can pause follicle development by inducing quiescence-associated secretory phenotype, a reversible senescent-like state, which may benefit fertility preservation.

Background: Increasing evidence suggests that the biological activity of trophoblasts and M1-type macrophages plays a crucial role in recurrent spontaneous abortion. However, detailed mechanistic studies on the intercellular communication between these two cells at the maternal-fetal interface are not clear.

Methods: In this study, extracellular vesicles (EVs) were first isolated from the supernatant of M1 macrophages induced by THP-1 cells (M1-EVs), identified by transmission electron microscopy, exosome immunofluorescence uptake, and western blotting, and characterized by mRNA sequencing to screen for specific target genes by mRNA profiling. CCK8 and western blotting experiments were used to investigate the effects of M1-EVs on trophoblast proliferation and autophagy. Subsequently, target genes MPPED2 and PI3K/AKT signaling pathway were found by bioinformatics analysis of raw mRNA sequencing results. Western blotting and CCK8 experiments were used to reveal the potential mechanisms by which MPPED2 in M1-EVs regulates trophoblast function.

Results: M1 macrophages induce inflammatory responses in the mother and fetus, and M1 macrophages inhibit trophoblast autophagy and proliferative capacity by secreting EVs. By mRNA transcriptome sequencing, MPPED2, among others, were identified as the most up-regulated mRNAs in M1-EVs-treated trophoblasts. Further functional experiments indicate that M1 macrophage-derived exosomes may regulate PI3K/AKT pathway activity by transferring MPPED2, leading to reduced autophagy and proliferation activity in trophoblasts.

Conclusion: Our findings suggest that MPPED2 from exosomes plays an important role in intercellular communication between M1 macrophages and the trophoblast, elucidating a novel mechanism by which M1 macrophages regulate trophoblast function and its role in recurrent spontaneous abortion.

The generation of haploid gametes is a hallmark of sexual reproduction achieved through a complex, albeit tightly regulated, reductional cell division known as meiosis. While the molecular underpinnings of meiosis have been extensively characterized in eutherian mammalian models, key aspects-particularly those governing chromosome synapsis and recombination-remain poorly understood in non-eutherian mammals and non-model vertebrates. This knowledge gap is especially relevant for understanding genome evolution, with a focus on sex chromosomes. Comparative studies across diverse vertebrate lineages are therefore essential to uncover conserved and lineage-specific features of meiotic regulation. In this review, we explore the evolutionary dynamics of meiosis in vertebrates, emphasizing how the meiotic program influences genome architecture and the evolution of heteromorphic sex chromosomes, especially the Y chromosome. As research in non-model species gains momentum, dissecting the diversity of meiotic mechanisms across taxa emerges as a key to understanding genome plasticity and evolutionary innovation.

Lactation is a cornerstone of mammalian life, providing nutrient-rich milk that supports offspring growth and development. Breastfeeding offers immediate and long-term benefits for infant and maternal health, shaping life-course health outcomes and reducing disease risks. Milk is a complex and dynamic fluid containing nutrients and bioactive molecules, and its composition is affected by environmental and genetic factors. Research on human milk falls into two primary areas: its biochemistry, which explores milk composition, and epidemiology, which examines the impact of lactation and breastfeeding on health. Together, these fields provide insights into the function of milk components. However, the genetic mechanisms that determine and influence milk composition remain poorly understood. Most findings linking genetics and milk composition have emerged from agricultural studies, where the mechanistic understanding is limited. In-vivo studies focus on mammary gland development, lactation, and cancer mechanisms, yet these interconnected topics are often divided across different research fields. Emerging evidence also suggests a role for genomic imprinting in the regulation of postnatal resource allocation via lactation and milk composition. This review aims to provide a summary of current knowledge on genetic influences affecting milk composition, offering a holistic perspective that unifies mammary gland physiology, milk biochemistry, lactation epidemiology, and genetics.

While maternal health has long been the focus of reproductive and developmental research, increasing attention is being given to the father's preconception health. Over the past two decades, growing evidence supports the Paternal Origins of Health and Disease (POHaD) paradigm, highlighting how paternal environmental factors prior to conception can significantly influence offspring development and long-term health. Rodent models have provided critical mechanistic insights into how paternal environmental exposure can alter the sperm epigenetic cargo. Emerging evidence indicates that paternal nutrition not only impacts the embryo but also the placenta. This review synthesizes current knowledge on how different types of paternal diet influence the sperm epigenome and the downstream consequences for the feto-placental unit and for pregnancy outcomes. We provide essential context: first, on the placenta's sensitivity to environmental influences, and second, on the variability across studies exploring paternal dietary exposures, highlighting key factors that may modulate paternal effects. We then examine how an altered diet affects the sperm epigenome and describe the physiological and molecular consequences on embryonic and placental development. Importantly, these modifications are linked to adverse outcomes including fetal growth restriction and placental insufficiency, and may therefore contribute to maternal complications such as preeclampsia, known for its long-term cardiovascular impact on women. Our review underscores the need to integrate paternal health into preconception care strategies, alongside maternal health, as it could significantly improve pregnancy outcomes and long-term health of both mothers and offspring.

Purpose: Purpose: To identify potential gut biomarkers linked to endometriosis for diagnosis and treatment.

Methods: The recruitment of this case-control study was done through the Endometriosis Health Profile-5 questionnaire, validated for endometriosis assessment. A total of 243 women completed the questionnaire and 73 women met the eligibility criteria of the study. Stool samples from the control group (n = 43) and patients with a positive diagnosis of endometriosis (n = 30) were collected and subjected to 16S ribosomal RNA (16S rRNA) gene sequencing using the V3-V4 regions. Various multivariate analysis approaches were used to assess diversity, composition, and abundance of intestinal microbiota.

Results: Among the 18 significantly different taxa (p < 0.05) between healthy controls and endometriosis patients, 3 families, 3 genera, and 12 species were identified. Endometriosis patients exhibited slightly higher diversity at the family and genus levels compared to controls (p > 0.05). This could indicate that endometriosis is characterized by the dominance of few species across diverse families and genera, associated with inflammation and estrogen signaling. Finally, several significant correlations (p < 0.05) were found between questionnaire variables related to "pain" and "infertility" items and certain families, genera, and species found in the endometriosis group.

Conclusion: The differential presence of estrobolome-presenting gut taxa between endometriosis patients and controls endorses the possible role of the gut microbiome in female reproductive health, offering potential microbial markers for endometriosis diagnosis, monitoring, and treatment. Identified taxa may serve as future prognostic, diagnostic, and therapeutic targets for diagnosis and personalized, preventive or palliative/curative treatment of endometriosis.

Summary sentence: Differential presence of estrobolome-related gut taxa in endometriosis (EMs) patients vs controls suggests a role of the gut microbiome in reproductive health, highlighting microbial markers for EMs diagnosis, monitoring, treatment, and future therapeutic targets.