scholarly journals Global, Survival, and Apoptotic Transcriptome during Mouse and Human Early Embryonic Development

2018 ◽  
Vol 2018 ◽  
pp. 1-16 ◽  
Author(s):  
D. Haouzi ◽  
I. Boumela ◽  
K. Chebli ◽  
S. Hamamah
Keyword(s):  
Embryonic Development ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Mouse Embryos ◽  
Signalling Pathways ◽  
Human Embryos ◽  
Special Focus ◽  
Mammalian Embryo ◽  
Human And Mouse ◽  
Early Human

Survival and cell death signals are crucial for mammalian embryo preimplantation development. However, the knowledge on the molecular mechanisms underlying their regulation is still limited. Mouse studies are widely used to understand preimplantation embryo development, but extrapolation of these results to humans is questionable. Therefore, we wanted to analyse the global expression profiles during early mouse and human development with a special focus on genes involved in the regulation of the apoptotic and survival pathways. We used DNA microarray technology to analyse the global gene expression profiles of preimplantation human and mouse embryos (metaphase II oocytes, embryos at the embryonic genome activation stage, and blastocysts). Components of the major apoptotic and survival signalling pathways were expressed during early human and mouse embryonic development; however, most expression profiles were species-specific. Particularly, the expression of genes encoding components and regulators of the apoptotic machinery were extremely stable in mouse embryos at all analysed stages, while it was more stage-specific in human embryos.CASP3,CASP9, andAIFwere the only apoptosis-related genes expressed in both species and at all studied stages. Moreover, numerous transcripts related to the apoptotic and survival pathway were reported for the first time such asCASP6andIL1RAPL1that were specific to MII oocytes;CASP2,ENDOG, andGFERto blastocysts in human. These findings open new perspectives for the characterization and understanding of the survival and apoptotic signalling pathways that control early human and mouse embryonic development.

scholarly journals Comparison of gene expression in fresh and frozen–thawed human preimplantation embryos

Reproduction ◽  
2012 ◽  
Vol 144 (5) ◽  
pp. 569-582 ◽  
Author(s):  
Lisa Shaw ◽  
Sharon F Sneddon ◽  
Daniel R Brison ◽  
Susan J Kimber
Keyword(s):  
Gene Expression ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Developmental Expression ◽  
Preimplantation Embryos ◽  
Human Embryos ◽  
Molecular Events ◽  
Reliable Source ◽  
Human Preimplantation Embryos ◽  
Early Human

Identification and characterisation of differentially regulated genes in preimplantation human embryonic development are required to improve embryo quality and pregnancy rates in IVF. In this study, we examined expression of a number of genes known to be critical for early development and compared expression profiles in individual preimplantation human embryos to establish any differences in gene expression in fresh compared to frozen–thawed embryos used routinely in IVF. We analysed expression of 19 genes by cDNA amplification followed by quantitative real-time PCR in a panel of 44 fresh and frozen–thawed human preimplantation embryos. Fresh embryos were obtained from surplus early cleavage stage embryos and frozen–thawed embryos from cryopreserved 2PN embryos. Our aim was to determine differences in gene expression between fresh and frozen–thawed human embryos, but we also identified differences in developmental expression patterns for particular genes. We show that overall gene expression among embryos of the same stage is highly variable and our results indicate that expression levels between groups did differ and differences in expression of individual genes was detected. Our results show that gene expression from frozen–thawed embryos is more consistent when compared with fresh, suggesting that cryopreserved embryos may represent a reliable source for studying the molecular events underpinning early human embryo development.

scholarly journals MicroRNA Expression Profiles Analysis in Sperm Reveals hsa-mir-191 is an Auspicious Omen of in Vitro Fertilization

2019 ◽  
Author(s):  
Hua Xu ◽  
Xin Wang ◽  
Zhikai Wang ◽  
Jianhui Li ◽  
Zhiming Xu ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryonic Development ◽  
Roc Curve ◽  
Small Rnas ◽  
Expression Profiles ◽  
High Quality ◽  
Vitro Fertilization ◽  
The Relationship ◽  
Early Human

Abstract Background: MicroRNAs (miRNAs) are a class of noncoding small RNAs that play important roles in many physiological processes by regulating gene expression. Previous studies have shown that the expression levels of total miRNAs increase during mouse embryonic development, and some miRNAs control the regulatory network in development progression. However, few studies have focused on the effects of miRNAs on early human embryonic development. The relationship between miRNAs and early human embryogenesis is still unknown. Results:In this study, RNA-seq data collected from sperm samples from 102 patients with a normal sperm index but treated with assisted reproductive technology (ART) were analyzed for the relationships between differentially expressed small RNAs and the fertilization rate (FR), blastocyst rate and high-quality embryo rate (HQER). The sperm samples with high hsa-mir-191 expression had a higher FR, effective embryo rate (EER) and HQER. hsa-mir-191 was used as a single indicator to predict the HQER. The receiver operating characteristic (ROC) curve had an area under the ROC curve (AUC) of 0.686. We also found that hsa-mir-191 expression is correlated with an abnormal sperm rate (cor = 0.29, p< 0.01). We also evaluated the relationship between hsa-mir-34c and early human embryo development in these 102 sperm samples and obtained negative results. Conclusions: These findings suggest that high hsa-mir-191-5p expression in sperm is associated with early human embryonic quality and that hsa-mir-191-5p could be used as a potential marker to screen high-quality sperm to improve the success rates of in vitro fertilization (IVF).

scholarly journals MicroRNA Expression Profiles Analysis in Sperm Reveals hsa-mir-191 is an Auspicious Omen of in Vitro Fertilization

2019 ◽  
Author(s):  
Hua Xu ◽  
Xin Wang ◽  
Zhikai Wang ◽  
Jianhui Li ◽  
Zhiming Xu ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Small Rna ◽  
Expression Profiles ◽  
Fertilization Rate ◽  
Small Rna Sequencing ◽  
High Quality ◽  
Potential Marker ◽  
Blastocyst Rate ◽  
The Relationship ◽  
Early Human

Abstract Background: MicroRNAs(miRNAs) are a class of non-coding small RNAs, which play important roles in many physiological processes by regulating gene expression. Previously studies show that the expression level of total miRNAs increased during the progression of mouse embryonic development, and some miRNAs control the regulatory network in the development progression. However, there are few studies focused on effects of miRNA on the early human embryonic development. The relationship of between miRNAs and early human embryogenesis is still unknown. Results In this study, 102 male sperm samples that were treated with assisted reproductive technology but with normal sperm index were collected for small RNA sequencing, and the relationships between differentially expressed small RNA and fertilization rate, blastocyst rate and high-quality embryo rate were analyzed. The results showed that the sperm samples with high expression of hsa-mir-191 had higher fertilization rate, blastocyst rate and high-quality embryo rate. Using hsa-mir-191 as a single indicator to predict high-quality embryo rate, the ROC curve has an AUC of 0.686. At the same time, we also found that the expression of hsa-mir-191 has a certain correlation with sperm abnormality rate (cor = 0.29, p< 0.01). In addition, we also evaluated the relationship between hsa-mir-34c and human early embryo development in these 102 sperm samples, and obtained negative results. Conclusions These findings suggest that higher hsa-mir-191-5p expression with better human early embryonic development, and indicate that the hsa-mir-191-5p could be a potential marker to access the quality of IVF.

De novo deletions of SNRPN exon 1 in early human and mouse embryos result in a paternal to maternal imprint switch

10.1038/75629 ◽  
2000 ◽  
Vol 25 (1) ◽  
pp. 74-78 ◽  
Author(s):  
Beata Bielinska ◽  
Susan M. Blaydes ◽  
Karin Buiting ◽  
Tao Yang ◽  
Malgorzata Krajewska-Walasek ◽  
...  
Keyword(s):  
De Novo ◽  
Mouse Embryos ◽  
Exon 1 ◽  
Human And Mouse ◽  
Early Human

scholarly journals Regulation of the cell cycle in early mammalian embryos and its clinical implications

2019 ◽  
Vol 63 (3-4-5) ◽  
pp. 113-122 ◽  
Author(s):  
Lenka Radonova ◽  
Tereza Svobodova ◽  
Martin Anger
Keyword(s):  
Cell Cycle ◽  
Embryonic Development ◽  
Rapid Development ◽  
Early Embryonic Development ◽  
Human Embryos ◽  
Valuable Insight ◽  
Mammalian Embryo ◽  
Early Mammalian Embryo ◽  
Mammalian Embryos ◽  
Cycle Monitoring

Early embryonic development is characterized by a plethora of very complex and simultaneously operating processes, which are constantly changing cellular morphology and behaviour. After fertilization, blastomeres of the newly created embryo undergo global epigenetic changes and simultaneously initiate transcription from the zygotic genome and differentiation forming separate cell lineages. Some of these mechanisms were extensively studied during the last several decades and valuable insight was gained into how these processes are regulated at the molecular level. We have, however, a still very limited understanding of how multiple events are coordinated during rapid development of an early mammalian embryo. In this review, we discuss some aspects of early embryonic development in mammals, namely the fidelity of chromosome segregation and occurrence of aneuploidy, as well as the clinical applications of cell cycle monitoring in human embryos.

scholarly journals Intrinsic Activation of β-catenin Signaling by CRISPR/Cas9-mediated Exon Skipping Contributes to Immune Evasion in Hepatocellular Carcinoma

2021 ◽  
Author(s):  
Masafumi Akasu ◽  
Shu Shimada ◽  
Ayano Kabashima ◽  
Yoshimitsu Akiyama ◽  
Masahiro Shimokawa ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Immune Evasion ◽  
Molecular Mechanisms ◽  
Nuclear Translocation ◽  
Expression Profiles ◽  
Gene Set Enrichment Analysis ◽  
Clinical Samples ◽  
Engineering System ◽  
Human And Mouse ◽  
Hcc Cells

Abstract Comprehensive analysis of clinical samples has recently identified molecular and immunological classification of hepatocellular carcinoma (HCC), and the CTNNB1 (β-catenin)-mutated subtype exhibits distinctive characteristics of immunosuppressive tumor microenvironment. For clarifying the molecular mechanisms, we first established human and mouse HCC cells with exon 3 skipping of β-catenin, which promoted nuclear translocation and activated the Wnt/β-catenin signaling pathway, by using newly developed multiplex CRISPR/Cas9-based genome engineering system. Gene set enrichment analysis indicated downregulation of immune-associated gene sets in the HCC cells with activated β-catenin signaling. T cell killing assays demonstrated that the mouse Ctnnb1Δex3 HCC cells evaded immune surveillance. Comparative analysis of gene expression profiles between HCC cells harboring wild-type and exon 3 skipping β-catenin elucidated that the expression levels of eight cytokines were commonly decreased in human and mouse β-catenin-mutated HCC cells. Public exome and transcriptome data of 373 human HCC samples showed significant downregulation of five candidate cytokine genes, CCL20, CXCL1, CXCL2, NAMPT and VEGFA, in HCC tumors with β-catenin hotspot mutations. Taken together, this study discovered that cytokine controlled by β-catenin signaling activation could contribute to immune evasion, and provided novel insights into cancer immunotherapy for the β-catenin-mutated HCC subtype.

61 Expression Profiles and Functional Roles of H3.3 and HIRA in Bovine Early Embryos

2018 ◽  
Vol 30 (1) ◽  
pp. 169
Author(s):  
K. Zhang ◽  
H. Wang
Keyword(s):  
Embryonic Development ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Mrna Level ◽  
Histone H3 ◽  
Specific Pattern ◽  
Early Embryonic Development ◽  
Early Embryos ◽  
Zygote Development

Early embryo death is one major reason for poor reproductive efficiency in dairy cows. In particular, ~20 to 50% of high-producing cows are subject to pregnancy loss during the first week of gestation, indicating the importance of embryonic development from fertilization to the blastocyst stage. To highlight this importance, multiple critical molecular and developmental events, including zygote reprogramming, maternal RNA decay, and embryonic genome activation, occur during bovine pre-implantation development. However, the molecular mechanisms of these events have yet to be defined. H3.3 is a histone H3 variant that encoded by 2 genes, namely, H3F3A and H3F3B. It is generally believed that H3.3 is closely related to active transcribed genes. Of interest, H3.3 required for establishing proper chromatin structure during mouse oogenesis. Immediately following fertilization, H3.3 is incorporated to parental chromatins and essential for blastocyst formation in mice. HIRA is a chaperone for H3.3 deposition and indispensable for zygote development. Previously, our results showed that H3.3 is needed for bovine early embryonic development. Herein, experiments were designed to determine the mechanisms of functional requirement of H3.3 in bovine early embryos. Slaughterhouse-derived cumulus–oocyte complexes (COC) were matured in vitro and IVF was performed. To knock down genes of interest, small interfering (si)RNAs were delivered into zygotes via microinjection. The qPCR results showed that H3F3A mRNA level is stable, whereas H3F3B and HIRA mRNA are dynamic during early embryonic development (4 replicates). The mRNA abundance of H3F3B is significantly higher than that of H3F3A (4 replicates; P < 0.05), which is also found in mouse and human. Immunostaining results revealed a stage-specific pattern for the localization of H3.3 in bovine early embryos, and the H3.3 signal was not different between paternal and maternal pronuclei in zygotes, which was different from the pattern in mice. The siRNA-mediated silencing of H3.3 dramatically reduces the expression of CTGF (a putative trophectoderm marker) in bovine blastocysts (3 replicates; P < 0.05). Furthermore, we found that the signal intensity of dimethylation of histone H3 lysine 36 (H3K36me2) and linker histone H1 decreases in H3.3-ablated embryos, which is similar to CHD1 knockdown (3 replicates; P < 0.05). However, no difference was found for the intensity of trimethylation of histone H3 lysine 4, dimethylation of histone H3 lysine 9 (H3K9me2) and splicing factor 3 B1 (SF3B1). We also found that HIRA deletion does not affect bovine early embryonic development. Taken together, the results described herein suggest that H3.3 is required for proper epigenetic modifications and H1 deposition during bovine early embryonic development. This project was supported by National Natural Science Foundation of China grant (No. 31672416) and the Fundamental Research Funds for the Central Universities.

scholarly journals Human and mouse embryonic development, metabolism and gene expression are altered by an ammonium gradient in vitro

Reproduction ◽  
2013 ◽  
Vol 146 (1) ◽  
pp. 49-61 ◽  
Author(s):  
D K Gardner ◽  
R Hamilton ◽  
B McCallie ◽  
W B Schoolcraft ◽  
M G Katz-Jaffe
Keyword(s):  
Gene Expression ◽  
Amino Acids ◽  
Transcriptome Analysis ◽  
Human Embryo ◽  
Culture Media ◽  
Expression Profiles ◽  
Human Embryos ◽  
Embryo Metabolism ◽  
Human And Mouse

Ammonium is generated in culture media by the spontaneous deamination of amino acids at 37 °C and through the metabolism of amino acids by human embryos. The appearance of ammonium is a time-dependent phenomenon and can compromise embryo physiology, development and viability. In this study, the effects of a gradient of ammonium on the development, metabolism and transcriptome of human and mouse embryos were investigated. Pronucleate oocytes were cultured in the presence of an ammonium gradient that mimicked the spontaneous deamination of Eagle's amino acids together with 1 mM glutamine. All embryos were cultured in sequential media G1/G2 at 5% O2, 6% CO2 and 89% N2. Human embryo metabolism was assessed through a non-invasive fluorometric analysis of pyruvate consumption. Transcriptome analysis was performed on the resultant blastocysts from both species using a microarray technology. Embryo development prior to compaction was negatively affected by the presence of low levels of ammonium in both species. Human embryo metabolism was significantly inhibited after just 24 and 48 h of culture. Transcriptome analysis of blastocysts from both species revealed significantly altered gene expression profiles, both decreased and increased. Functional annotation of the altered genes revealed the following over represented biological processes: metabolism, cell growth and/or maintenance, transcription, cell communication, transport, development and transcription regulation. These data emphasize the enhanced sensitivity of the cleavage-stage embryo to its environment and highlight the requirement to renew culture media at frequent intervals in order to alleviate the in vitro induced effects of ammonium build-up in the environment surrounding the embryo.

scholarly journals Pleiotropic roles of metallothioneins as regulators of chondrocyte apoptosis and catabolic and anabolic pathways during osteoarthritis pathogenesis

2016 ◽  
Vol 75 (11) ◽  
pp. 2045-2052 ◽  
Author(s):  
Yoonkyung Won ◽  
Youngnim Shin ◽  
Churl-Hong Chun ◽  
Yongsik Cho ◽  
Chul-Won Ha ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Hypoxia Inducible Factor ◽  
Gene Expression Profiles ◽  
Cartilage Destruction ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Chondrocyte Apoptosis ◽  
Double Knockout ◽  
Nicotinamide Phosphoribosyltransferase ◽  
Human And Mouse

ObjectiveThe zinc-ZIP8-MTF1 axis induces metallothionein (MT) expression and is a catabolic regulator of experimental osteoarthritis (OA) in mice. The main aim of the current study was to explore the roles and underlying molecular mechanisms of MTs in OA pathogenesis.MethodsExperimental OA in mice was induced by destabilisation of the medial meniscus or intra-articular injection of adenovirus carrying a target gene (Ad-Zip8, Ad-Mtf1, Ad-Epas1, Ad-Nampt, Ad-Mt1 or Ad-Mt2) into wild type, Zip8fl/fl; Col2a1-Cre, Mtf1fl/fl; Col2a1-Cre and Mt1/Mt2 double knockout mice. Primary cultured mouse chondrocytes were infected with Ad-Mt1 or Ad-Mt2, and gene expression profiles analysed via microarray and reverse transcription-PCR. Proteins in human and mouse OA cartilage were identified via immunostaining. Chondrocyte apoptosis in OA cartilage was determined using terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick end labelling (TUNEL).ResultsMTs were highly expressed in human and mouse OA cartilage. Hypoxia-inducible factor 2α, nicotinamide phosphoribosyltransferase and several proinflammatory cytokine pathways, as well as the zinc-ZIP8-MTF1 axis were identified as upstream regulators of MT expression. Genetic deletion of Mt1 and Mt2 enhanced cartilage destruction through increasing chondrocyte apoptosis. Unexpectedly, aberrant overexpression of MT2, but not MT1, induced upregulation of matrix-degrading enzymes and downregulation of matrix molecules through nuclear factor-kappa B (NF-κB) and activator protein-1 (AP-1) activation, ultimately leading to OA.ConclusionsMTs play an antiapoptotic role in post-traumatic OA. However, aberrant and chronic upregulation of MT2 triggers an imbalance between chondrocyte anabolism and catabolism, consequently accelerating OA development. Our findings collectively highlight pleiotropic roles of MTs as regulators of chondrocyte apoptosis as well as catabolic and anabolic pathways during OA pathogenesis.

scholarly journals Unraveling the Spatiotemporal Human Pluripotency in Embryonic Development

2021 ◽  
Vol 9 ◽  
Author(s):  
Daniela Ávila-González ◽  
Wendy Portillo ◽  
Guadalupe García-López ◽  
Anayansi Molina-Hernández ◽  
Néstor E. Díaz-Martínez ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Molecular Mechanisms ◽  
Primordial Germ Cells ◽  
Mouse Embryos ◽  
Human Embryos ◽  
Self Organized ◽  
Intermediate States ◽  
Human Embryogenesis ◽  
Signaling Center

There have been significant advances in understanding human embryogenesis using human pluripotent stem cells (hPSCs) in conventional monolayer and 3D self-organized cultures. Thus, in vitro models have contributed to elucidate the molecular mechanisms for specification and differentiation during development. However, the molecular and functional spectrum of human pluripotency (i.e., intermediate states, pluripotency subtypes and regionalization) is still not fully understood. This review describes the mechanisms that establish and maintain pluripotency in human embryos and their differences with mouse embryos. Further, it describes a new pluripotent state representing a transition between naïve and primed pluripotency. This review also presents the data that divide pluripotency into substates expressing epiblast regionalization and amnion specification as well as primordial germ cells in primates. Finally, this work analyzes the amnion’s relevance as an “signaling center” for regionalization before the onset of gastrulation.

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