Biology of Reproduction最新文献

RTCB (RNA 2',3'-cyclic phosphate and 5'-OH ligase) is a unique 3'-5' RNA ligase with diverse physiological functions in metazoans. To further explore the role of RTCB in reproduction, we generated a Rtcb conditional knockout mouse model using the Ddx4-Cre system. The complete absence of viable Rtcb-/- offspring indicated embryonic lethality. Integrated analyses of histology, immunohistochemistry, quantitative PCR, and published single-cell RNA sequencing (scRNA-seq) data revealed that Rtcb is expressed throughout embryogenesis, with pronounced upregulation during gastrulation, and that Rtcb knockout resulted in gastrulation failure around embryonic day 6.5 (E6.5), which was accompanied by decreased proliferation and increased apoptosis of embryonic cells. Notably, NUSAP1 (nucleolar and spindle-associated protein 1) is vital for gastrulation as it participates in cell division. In NIH 3T3 cells, knockdown of Rtcb led to destabilization of Nusap1 mRNA, suggesting that NUSAP1 might function downstream of RTCB. Moreover, Rtcb is a target gene of YY1 (Yin-Yang-1), a transcription factor crucial for gastrulation. Expression profiling through scRNA-seq revealed spatiotemporal coordination among Yy1, Rtcb, and Nusap1 between E3.5 and E7.5. Collectively, our findings demonstrate that RTCB is essential for early mouse embryogenesis and propose the presence of a YY1-RTCB-NUSAP1 axis that maintains proper cell proliferation for successful gastrulation.

In each wave, only a small portion of primordial follicles are selectively activated into the growing follicular pool, and the majority of primordial follicles remain in a relatively quiescent state. Hypoxanthine is present in the follicular fluid with high concentrations and inhibits phosphodiesterase (PDE) activity. $$ contribute to the promotion of the cyclic activation of primordial follicles.

In this study, we utilized single-nucleus RNA sequencing to quantify alterations in the gene expression programs of mouse placentaeconceived through in vitro fertilization (IVF). We identified genetic programs exhibiting both global and cell type-specific differences between IVF and natural in vivo fertilization groups. Gene set enrichment analysis revealed pathways associated with parietal trophoblast giant cell differentiation and implicated in the regulation of lactation (placental lactogens), along with hypoxia-inducible factor-dependent gene expression. Importantly, IVF-derived conceptuses showed increased abortion rates when their surrogate mothers were exposed to hypoxia (10.5% O2) during pregnancy (from E7.5 to 12.5). Collectively, our findings shed light on the cellular and molecular underpinnings driving differences in pregnancy outcomes associated with these conception methods and indicate that IVF can sensitize embryos to additional stressful events, as proposed by the developmental origin of health and disease hypothesis.

Adenosine (ADO), a purinergic system ligand, plays important roles in several physiological processes, including proliferation, differentiation, immunity, development, and reproduction. The activation of various intracellular signaling pathways by ADO is mediated through ADO receptors, ADORA1, ADORA2A, ADORA2B, and ADORA3. Although the importance of ADO during pregnancy has been studied in some species, the expression of ADO receptors and the roles of ADO at the maternal-conceptus interface have not been studied in pigs. Therefore, we investigated the expression and regulation of ADO receptors and the function of ADO at the maternal-conceptus interface in pigs. The expression of ADO receptors in the endometrium changed dynamically during pregnancy, and the levels of ADORA1, ADORA2A, and ADORA3 expression were greater during early pregnancy than during the estrous cycle. During pregnancy, both conceptus and chorioallantoic tissues expressed ADO receptors. Estradiol-17β, interleukin-1β, and/or interferon-γ increased the endometrial expression of ADORA1 and ADORA2A. Total recoverable amounts of ADO in the uterine lumen were greater on Day 15 of pregnancy than on Day 15 of the estrous cycle. Increasing doses of ADO increased the expression of prostaglandin (PG) synthetic enzymes, PGF2α secretion, and the activation of mitogen-activated protein kinases in endometrial epithelial (pUE) cells, and migration of conceptus trophectoderm cells in vitro. These data suggest that conceptus-derived factors affect the endometrial expression of ADO receptors and ADO production. Therefore, ADO plays important roles in the regulation of pUE cells and conceptus trophectoderm cell functions at the maternal-conceptus interface to establish and maintain pregnancy in pigs.

Placental cotyledons play a crucial role in material exchange and gas transportation between the ruminant mother and fetus and their proper development is essential for the intrauterine growth and development of the fetus. This study aimed to investigate the relationships between cotyledon size, litter size, and birth weight, while also identifying the key candidate genes involved in cotyledon development. Based on 95 postpartum goat placentas, significant positive associations were observed between cotyledon area, litter size, and birth litter weight. Histological examination showed a significant increase in capillary density with increasing cotyledon area, accompanied by significant changes in the cotyledon villus structure. Through the RNA-seq analysis of large (diameter > 30 mm), medium (diameter between 10-30 mm), and small (diameter < 10 mm) placental cotyledons, a total of 1103 differentially expressed genes (DEGs) were identified while RT-qPCR experiments confirmed the reliability of the sequencing results. The KEGG analysis indicated that the identified DEGs were primarily enriched in pathways such as the PI3K-AKT signaling pathway, the ECM-receptor interaction, the JAK-STAT signaling pathway, and the Estrogen signaling pathway. In conjunction with Gene Ontology functional annotations and Gene Set Enrichment Analysis (GSEA), the Spp1, Efna3, Igta3, and Csf1r genes were identified as potential candidate genes affecting the development and angiogenesis of goat placental cotyledons. These findings provide important insights into the mechanisms underlying the development of goat cotyledons and their relationship with the reproductive performance of goats.

Fertility is a significant determinant of productivity in the beef cattle industry and young bulls are often fed highly caloric diets to accelerate growth. However, the effects of such nutritional strategies on early embryonic development remain poorly described. We hypothesized that feeding young bulls a high-gain diet would compromise early embryonic development in vitro. Twenty-nine bulls (13 months of age) were fed a moderate (target average daily gain = 1.22 kg/day; n = 15) or high gain (target average daily gain = 1.81 kg/day; n = 14) diet for 114 days. The high-gain diet increased average daily gain, final body weight, and subcutaneous backfat thickness. Semen was collected and frozen from each bull after 114 d of treatment and used for in vitro fertilization. Semen from high-gain bulls reduced embryo cleavage and early blastocyst formation. Blastocysts from high-gain bulls had fewer blastomeres, altered inner cell mass to trophectoderm ratio, and increased apoptosis. In addition, time-lapse embryo culture indicated that semen from high-gain bulls tended to reduce embryo cleavage and development to morula and early blastocyst stages, and delayed temporal progression to morula and blastocyst hatching. Similarly, semen from high-gain bulls increased cytoplasmic fragmentation of blastomeres during the first three cleavage events. These findings indicate that overnutrition of young bulls produces semen that compromises embryo development and quality, highlighting the need to develop feeding strategies to optimize growth and reproductive potential in cattle. Overnutrition of young, growing bulls results in semen that delays embryonic development and decreases embryo quality.

An optimal uterine luminal fluid (ULF) microenvironment is essential for successful pregnancy establishment and progression. Despite this, the specific factors influencing early embryo and conceptus (embryo and extra-embryonic structures) survival remain poorly understood. Lipid moieties dominate the ULF metabolome and are thought to play crucial roles in embryo and conceptus development. As such, we aimed to comprehensively profile the bovine ULF lipidome at four key times of the estrous cycle-Days 0, 7, 10, and 14. Our analysis identified 80 distinct lipids, which were categorized into 29 metabolic pathways. We observed significant (P ≤ 0.05) temporal variation in lipid abundance, with certain lipids uniquely present on specific days. Notably, carnitine derivatives exhibited pronounced changes in abundance throughout the cycle. These findings highlight the dynamic nature of ULF lipid composition and its potential influence on early reproductive events. We also discuss the implications of these results for enhancing both in vivo and in vitro embryo development.

The aryl hydrocarbon receptor (AHR) plays a role in the development, function, and xenobiotic metabolism in multiple tissues, including the ovary. The AHR is a member of the Per/Arnt/Sim basic helix-loop-helix (bHLH) family of transcription factors. Endogenous and exogenous ligands activate the AHR signaling pathway. Activation of the AHR pathway leads to transcription of downstream targets. This review highlights the involvement that the AHR has in ovarian development and function. This review focuses on the involvement that the AHR has in both folliculogenesis and steroidogenesis, including the hypothalamic-pituitary-gonadal axis. Several studies indicate that the AHR mediates normal ovarian function and can influence xenobiotic metabolism in the ovary. Further, the review summarizes the involvement of the AHR in ovarian diseases such as infertility, polycystic ovarian syndrome, and ovarian cancer, and the potential for targeting the AHR as a therapeutic option for ovarian disease. Finally, this review addresses gaps in the literature that can be addressed to further the understanding of the role that the AHR has in the ovary and how its role can be leveraged in the clinic. This review examines the role that the aryl hydrocarbon receptor (AHR) plays in ovarian function (folliculogenesis, steroidogenesis, and xenobiotic metabolism) and disease.

Placental complications resulting in fetal growth restriction have been associated with dysregulated placental gene expression tied to an increased risk of schizophrenia. In rat offspring, it has been demonstrated that ∆9-tetrahydrocannabinol exposure in pregnancy results in fetal growth restriction and schizophrenia-like phenotypes (e.g., decreased pre-pulse inhibition of the acoustic startle response). However, it remains elusive if prenatal ∆9-tetrahydrocannabinol exposure induces this schizophrenia signature of placental gene expression. Therefore, our objective was to determine if these established predictive markers of schizophrenia are altered in a preclinical model of gestational oral ∆9-tetrahydrocannabinol exposure in rodents. We observed significantly reduced fetal weights in male and female prenatal ∆9-tetrahydrocannabinol-exposed offspring in the absence of maternal pregnancy outcomes. Placentae from ∆9-tetrahydrocannabinol-exposed males and females revealed altered expression of genes previously identified in human transcriptomic datasets of schizophrenia (i.e., Furin, Rccd1, and Atp5mk), with some expression changes being sex-specific (i.e., Eif5, Rps10, Vps33b, and Iqgap1). A subset of these genes were found differentially expressed in human BeWo cells exposed to ∆9-tetrahydrocannabinol. Targets were next examined in the adult rodent (postnatal day70) brain, and a subgroup of these genes (i.e., Furin, Rps10, and Rccd1) were increased concomitant with schizophrenia-like behavior (e.g., decreased pre-pulse inhibition). We further detected ∆9-tetrahydrocannabinol-induced upregulation of FURIN in patient-derived cerebral organoids, an effect observed in both control and schizophrenia cell lines. Collectively, these findings demonstrate prenatal ∆9-tetrahydrocannabinol exposure can lead to altered gene expression in established prioritized markers of schizophrenia in the placenta in both animal and human models.

Trophoblast stem cells (TSCs) serve as a critical model for understanding placental development, early embryo-maternal interactions, and pregnancy establishment in mammals. In cattle, the developing trophectoderm plays an essential role in conceptus elongation and secretion of factors necessary for maternal recognition of pregnancy. Building on previous work identifying signaling pathways regulating bovine TSC self-renewal and differentiation, we report the generation and characterization of transformed bovine TSC (bTSC) lines derived from blastocysts via lentiviral transduction of simian vacuolating virus 40 large T antigen. These rapidly proliferating TSC cell lines, maintained in the presence of Rho-associated protein kinase (ROCK) inhibition, retain key morphological and transcriptional characteristics of bovine TSCs. Upon transforming growth factor β-induced differentiation, they exhibit morphological and molecular changes consistent with trophoblast maturation. To evaluate their utility for functional studies, we demonstrated stable gene introduction of tdTomato and EGFP using lentiviral vectors and employed CRISPR/Cas9-mediated gene editing to target lentiviral EGFP integration sites, confirming efficient gene deletion. Additionally, proteomic analysis of conditioned medium identified secreted proteins with potential roles in embryo-uterine interactions, aligning with factors previously reported in bovine conceptus secretomes. These findings establish transformed bTSC lines as a valuable model for investigating bovine trophoblast biology, functional gene studies, and trophoblast-endometrial signaling. By providing a renewable in vitro system with stable proliferative capacity, these cell lines enable further exploration of the molecular mechanisms governing early pregnancy in cattle.