Molecular Reproduction and Development最新文献

Recurrent pregnancy loss (RPL) is one of obstetrical diseases with no effective therapy methods. Trophoblast cell dysfunction and inflammation induce embryo implantation insufficiency, thereby resulting in RPL. OTU deubiquitinase with linear linkage specificity (OTULIN) plays a role in regulating the immune response and cell death. However, the role of OTULIN in RPL remains unclear. Spontaneous abortion mouse model and lipopolysaccharide-treated HTR-8/SVneo cells were used to investigate the role of OTULIN in RPL. OTULIN expression was downregulated in the labyrinth trophoblast of RPL mice and LPS-treated trophoblast cells. The embryonic reabsorption rate was decreased in OTULIN-overexpressed spontaneous abortion mice, accompanied with the increase in placental/fetus weight ratio. OTULIN overexpression significantly inhibited apoptosis in vivo and in vitro, as evidenced by the decrease in the activity of caspase 3. The expression of pro-inflammatory cytokines was decreased with OTULIN overexpression. Moreover, OTULIN overexpression decreased p-IκBα/IκBα and p-p65/p65 ratio. The nuclear translocation of NF-κB was suppressed via OTULIN overexpression both in vivo and in vitro. Our study suggested that OTULIN deficiency might cause inflammation and trophoblast abnormalities in RPL. The supplementation with OTULIN might alleviate the development of RPL via inhibiting NF-κB mediated inflammation response.

Hypoxia-inducible factor 1-alpha (HIF-1α) is essential for glycolysis regulation. Its expression in the endometrium is significantly reduced in recurrent implantation failure (RIF), indicating that lower levels of HIF-1α may contribute to embryo implantation failure. O-GlcNAcylation is a dynamic posttranslational modification mediated by O-GlcNAc transferase (OGT), known to regulate HIF-1α in cancer cells. However, it remains unclear whether OGT affects glycolytic processes in uterine endometrial stromal cells (ESCs) and its potential role in embryo implantation. This study utilized In Vitro and In Vivo experiments to investigate the role of OGT in decidualization and embryo implantation, along with its underlying mechanisms. Our findings show that OGT expression is significantly reduced in the endometrium of patients with RIF. Additionally, OGT knockdown led to failed embryo implantation in mice. Further analysis revealed that OGT promotes decidualization by stabilizing HIF-1α, which enhances glycolytic activity. Inhibiting OGT resulted in insufficient decidualization among human ESCs. Moreover, our results indicate that OGT partially regulates CCL2 secretion by maintaining HIF-1α levels within human ESCs, which is essential for successful embryo implantation. Based on these findings, we propose that OGT represents a novel and promising therapeutic target for both the diagnosis and treatment of RIF.

In this study, we examine the transcriptional profiles of eight stages of preimplantation embryo development, from germinal vesicle (GV) oocytes to blastocysts, in in vitro fertilized sheep embryos using RNA sequencing. Between 12,744 and 15,539 genes were detected from the GV moving forward to blastocyst, with cluster analysis revealing two distinct clusters related to developmental stage. The largest change for differentially expressed genes (DEGs) numbers took place between 2- to 4-cell comparison and 4- to 8-cell comparison, with a c. 35-fold increase from 19 to 670 DEGs. This suggests that zygotic genome activation occurred at 8-cell stage in sheep. All the DEGs had gene ontology annotations in three classes, and these comprised 53 subclasses. Most of the pathways (89 of 93) were enriched through KEGG analysis of the DEGs, and the most enriched pathway was oxidative phosphorylation. The 2- and 16-cell stage embryos possessed the highest and lowest numbers, respectively, of single nucleotide polymorphisms and insertion-deletion markers. Five major types of alternative splicing were found. A total of 2361 transcription factor genes were mutually expressed among the eight developmental stages. The top five differential transcription factor protein families were Zf-C2H2, bHLH, Homeobox, Others, and HMG. This is the first study to investigate the transcriptional profiling of sheep preimplantation embryos at different developmental stages, and it displays a comprehensive transcriptome landscape in sheep oocytes and embryos.

The functional role of estrogen receptor activation factor (E-RAF), a 66 kDa protein in mediating the biological actions of estrogen and progesterone in goat uterus, has been highlighted in this work. Here we report that E-RAF stimulates the expression of splicing factor gene, arginine/serine rich in goat uterine nuclei (GenBank Accession No. MT473439). cDNA subtractive hybridization method using nuclear run-on transcription assay was used to identify the differentially expressed genes. The upregulation of the splicing factor gene was demonstrated further using a PCR-based nuclear run-on transcription system. Confocal microscopic analysis has confirmed the endoplasmic reticulum as the primary site of intracellular location of E-RAF. The effect of estrogen and progesterone on the intracellular distribution of E-RAF and the molecular mechanisms that underlie the transport of E-RAF to nucleus in the presence of progesterone has been studied. Our results offer a new perspective in steroid hormone action in uterus.

This study focused on exploring the mechanism of transcription factor FOXD3 promoting the proliferation, migration, and invasion of trophoblast through ELAV1/TLR4 axis. The placenta villi from pregnancy patients with antiphospholipid syndrome (APS) and pregnancy controls were collected, along with the HTR-8/SVneo cell lines were obtained to detect the FOXD3, ELAV1, and TLR4 expressions using qRT-PCR and western blot. The interaction of ELAV1 with TLR4 mRNA was verified using RNA immunoprecipitation. The binding of FOXD3 with ELAV1 was detected using Chromatin Immunoprecipitation and dual luciferase reporter gene assay. After cell transfection, the cell proliferation, cell cycle distribution, invasion, and migration of the HTR-8/SVneo cell line were also measured. FOXD3, ELAV1, and TLR4 were elevated in the placenta villi of APS patients. TLR4 knockdown can promote the proliferation, invasion, and migration ability of HTR-8/SVneo cells. ELAV1 can bind TLR4 mRNA and increase its stability. TLR4 overexpression can inhibit the promotive effect of ELAV1 knockdown on HTR-8/SVneo cell biological functions. FOXD3 can bind the ELAV1 promoter and increase its transcription level to mediate HTR-8/SVneo cell biological functions. FOXD3 can bind and increase ELAV1 expression to stabilize TLR4 mRNA level, thereby increasing TLR4 expression and inhibiting the proliferation, invasion, and migration ability of trophoblast.

Ovarian tumor ubiquitinating 6A (OTUD6A) is a deubiquitinating enzyme whose aberrant expression has been linked to various diseases, including inflammation and prostate cancer. Research indicates that deubiquitinating enzymes (DUBs) play a significant role in spermatogenesis in mice. However, the role of OTUD6A in spermatogenesis remains unclear. To investigate the function of OTUD6A in mouse spermatogenesis, we generated Otud6a-knockout mice using the CRISPR/Cas9 system. Our results showed that OTUD6A is predominantly expressed in the testis and localized to the cytoplasm of spermatogonia and spermatocytes. Although no significant differences were observed in testicular size or morphology between Otud6a-knockout and wild-type mice, the knockout mice exhibited increased germ cell apoptosis, decreased epididymal sperm counts, abnormalities in sperm motility and subfertility. These findings indicate that Otud6a-knockout leads to male subfertility in mice.