Transcriptomic analysis of Ywhaz knockout in preimplantation mouse embryos
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE294735 | GEO | 2025/08/31
REPOSITORIES: GEO
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Project description:Transcriptomic analysis of Ywhaz knockout in preimplantation mouse embryos
| PRJNA1251527 | ENA
Project description:To investigate the importance of DNA damage checkpoint regulation in mammalian preimplantation embryos, we overexpressed wild type and mutant form of CHK1 kinase in mouse zygotes and performed high-throughput RNA-sequencing of zygotes, late 2-cell embryos and blastocysts. Transcriptome analysis revealed that zygotes are particular sensitive to the perturbation of Chk1 governed DNA damage checkpoint. However, once zygotes can overcome DNA damage checkpoint induced cell cycle arrest, they can still have normal development.
2021-05-04 | GSE163765 | GEO
Project description:We systematically analyzed transcriptome in individual human oocytes and preimplantation embryos with a new single-cell RNA seq method we recently developed. We sequenced individual single cells or embryos at seven consecutive stages (mature oocytes, zygotes, 2-cell, 4-cell, 8-cell embryos, morulae, and blastocysts) and single hESC using single-cell RNA seq.
2016-05-13 | E-GEOD-71318 | biostudies-arrayexpress
Project description:Age of the donor animal has a significant influence on the oocyteM-bM-^@M-^Ys ability to complete maturation and acquire the mRNA and proteins required for normal embryonic development. It is well known that developmental competence is reduced in oocytes derived from juvenile animals when compared to that derived from adults. However, the molecular mechanisms associated with these differences are not well elucidated. The aim of this study was to analyze the transcriptome differences between adult versus prepubertal derived preimplantation embryos and to identify genes associated with oocyte developmental potential. We performed cDNA microarray analysis on populations of preimplantation embryos (8- to 16-cell and Day 7 blastocysts) derived from adult versus prepubertal Japanese Black cattle. Total RNA was extracted and amplified in a linear fashion and then subjected to the microarray hybridization using the Affymetrix GeneChip Bovine Genome Array. The Bovine Array contains 24 072 probe sets representing over 23000 transcripts and 19000 UniGene clusters. Following normalization of the microarray data, analysis revealed differences in gene expression abundance in 8- to 16-cell and blastocysts harvested from adult versus prepubertal heifers. Results indicated that a total of 591 and 490 genes were differentially expressed (M-bM-^IM-%2-fold) in 8- to 16-cell and blastocyst respectively between the two groups (p<0.01). In 8- to 16-cell stage, 373 genes were up-regulated and 218 gens down-regulated in adult when compared to the prepubertal heifer group. In case of blastocyst stage embryos, 242 genes were up-regulated and 248 genes were down-regulated in adult when compared to prepubertal group. The ovaries of adult cows (Japanese black cattle) were collected from local abattoir while ovaries of prepubertal Japanese black heifers (9 to 12 months old) were collected by spay device at several commercial farms. The ovaries were transported to the research laboratory in 0.67% (w/v) NaCl solution containing 100 mg/L kanamycin sulfate. COC were placed in 200 M-BM-5L drop of the maturation medium (TCM 199 + 10% FBS) and cultured at 38.5M-BM-0C in a humidified atmosphere of 5% CO2 in air for 20 to 22 h for in vitro maturation (IVM). After 20 to 22 h IVM, the COC were fertilized in vitro with frozenM-bM-^@M-^Sthawed semen of Japanese Black bull in BO medium. Six hours after IVF, oocytes were denuded of surrounding cumulus cells by gentle pipetting in a solution of 0.1% (w/v) hyaluronidase. The presumptive zygotes were cultured in CR1aa medium supplemented with fatty acid-free BSA (Sigma) for 2 days and cultured in CR1aa medium supplemented with 10% (v/v) fetal bovine serum (FBS) for next 5 days at 38.5 M-BM-0C in 5% CO2, 5% O2 and 90% N2 with high humidity (15 to 20 zygotes per 50 M-BM-5l microdrop). 8- to 16-cell refers to day 3, 70 to 72 hours post insemination( hpi) preimplantation embryo stage containing 8 to 16 cells, while blastocyst refer to pre-hatching day 7 expanded blastocysts. Pools of 8- to 16- cell embryos (50 in each sample) and blastocysts (10 in each sample) from prepubertal and adult groups were used for cDNA microarray.
2012-05-31 | E-GEOD-24596 | biostudies-arrayexpress
Project description:Clinically-discarded oocytes and preimplantation embryos at various stages were collected to perform proteome analysis.
2025-05-26 | PXD042633 | Pride
Project description:Metastasis is a major cause leading to mortality for lung cancer patients. We identified YWHAZ as a potential metastasis-promoting candidate and found that overexpression of YWHAZ promotes lung cancer cell proliferation, anchorage-independent growth, migration, and invasion in vitro, as well as tumorigenesis and metastasis in vivo. It not only increases cell protrusions and branchings but also induces epithelial-mesenchymal transition. Most importantly, YWHAZ protein could prevent £]-catenin from ubiquitination via its association with £]-catenin and enhance slug transcriptional activity which is regulated by £]-catenin/TCF signaling pathway. Moreover, YWHAZ expression was higher in tumors than in adjacent normal tissues in 63 Non-small-cell lung cancer (NSCLC) patients. NSCLC patients with high YWHAZ expressing tumors had shorter overall survival than those with low-expressing tumors. We conclude that YWHAZ play a critical role in promoting NSCLC metastasis. In this investigation, we used a lung cancer invasion cell model to identify the genes involved in cancer progression. YWHAZ is a potential oncogene whose expression is correlated to the survival of patients with breast, prostate and liver cancers. However, the role of YWHAZ in lung caner progression has not been reported, particularly in metastasis. Here, YWHAZ was ectopically expressed in lower invasive lung cancer cell line its impact on colonogenesis, migration and invasiveness was assessed. The underlying mechanism was explored by YWHAZ-expressed transfectants and microarrays and the clinical relevance was evaluated by quantitative RT-PCR.
2012-11-07 | E-GEOD-20318 | biostudies-arrayexpress
Project description:Maternal nutrition exclusively during the periconceptional period can induce remarkable effects on both oocyte maturation and early embryo development, which in turn can have lifelong consequences. The objective of this study was to evaluate the effect of maternal methionine supplementation on the transcriptome of bovine preimplantation embryos. Holstein cows were randomly assigned to one of two treatments differing in level of dietary methionine (1.89 Met vs. 2.43 Met % of metabolizable protein) from calving until embryo flushing. High quality preimplantation embryos from individual cows were pooled and then analyzed by RNA sequencing. A total of eight Holstein dairy cows were used in this study. Preimplantation embryos recovered from each cow were pooled in order to generate two replicates per cow assayed. Each pool consisted of 2-4 expanded blastocysts with excellent quality. Overall, a total of 16 embryo pools underwent RNA extraction, amplification, and subsequent sequencing.
2013-06-21 | E-GEOD-48147 | biostudies-arrayexpress
Project description:40-PE Smart-Seq of RNA isolated from single 2-cell embryos, single morulae, single blastocysts, single E7 embryos epiblast halves and single E7 trophectoderm halves. For all preimplantation stages, embryos were grown ex-vivo from three experimental groups of zygotes: zygotes injected with the mRNA of a recombinant OGA (named BtGH84) (Btgh_injected), zygotes injected with a catalytically dead version of BtGH84 (dBtgh_injected) (control nr.1) and non-injected zygotes (non_injected) (control nr.2). Postimplantation E7 embryos were grown from two experimental group of zygotes: Btgh-injected and dBtgh-injected (control) and transferred at the 2-cell stage to foster mothers. All samples from the same embryonic stage were processed for single embryo (or single-embryo single-tissue) Smart-Seq library preparation at the same time and then sequenced in the same NextSeq500 run. The different embryonic stages were processed on different days and sequenced in different NextSeq500 runs. Together with the raw fastq files, the table of gene counts obtained with featureCounts is provided, containing all samples that passed filtering steps and were input to the PCA and differential expression analysis (last column of Table S3 of the manuscript).
2024-03-30 | E-MTAB-12981 | biostudies-arrayexpress
Project description:Polycomb repressive complexes 1 and 2 (PRC1/2) maintain transcriptional silencing of developmental genes largely by catalyzing mono-ubiquitination of histone H2A at lysine 119 (H2AK119ub1) and trimethylation of histone H3 at lysine 27 (H3K27me3), respectively. How Polycomb domains are reprogrammed during mammalian preimplantation development remains largely unclear. Here we show that, although H2AK119ub1 and H3K27me3 are highly colocalized in gametes, they undergo differential reprogramming dynamics following fertilization. H3K27me3 maintains thousands of maternally biased domains up to the blastocyst stage, whereas maternally biased H2AK119ub1 distribution in zygotes is largely equalized at the two-cell stage. Notably, while maternal PRC2 depletion has a limited effect on global H2AK119ub1 in early embryos, it disrupts allelic H2AK119ub1 at H3K27me3 imprinting loci including Xist. By contrast, acute H2AK119ub1 depletion in zygotes does not affect H3K27me3 imprinting maintenance, at least by the four-cell stage. Importantly, loss of H2AK119ub1, but not H3K27me3, causes premature activation of developmental genes during zygotic genome activation (ZGA) and subsequent embryonic arrest. Thus, our study reveals distinct dynamics and functions of H3K27me3 and H2AK119ub1 in mouse preimplantation embryos.
2021-03-15 | GSE153529 | GEO
Project description:Polycomb repressive complexes 1 and 2 (PRC1/2) maintain transcriptional silencing of developmental genes largely by catalyzing mono-ubiquitination of histone H2A at lysine 119 (H2AK119ub1) and trimethylation of histone H3 at lysine 27 (H3K27me3), respectively. How Polycomb domains are reprogrammed during mammalian preimplantation development remains largely unclear. Here we show that, although H2AK119ub1 and H3K27me3 are highly colocalized in gametes, they undergo differential reprogramming dynamics following fertilization. H3K27me3 maintains thousands of maternally biased domains up to the blastocyst stage, whereas maternally biased H2AK119ub1 distribution in zygotes is largely equalized at the two-cell stage. Notably, while maternal PRC2 depletion has a limited effect on global H2AK119ub1 in early embryos, it disrupts allelic H2AK119ub1 at H3K27me3 imprinting loci including Xist. By contrast, acute H2AK119ub1 depletion in zygotes does not affect H3K27me3 imprinting maintenance, at least by the four-cell stage. Importantly, loss of H2AK119ub1, but not H3K27me3, causes premature activation of developmental genes during zygotic genome activation (ZGA) and subsequent embryonic arrest. Thus, our study reveals distinct dynamics and functions of H3K27me3 and H2AK119ub1 in mouse preimplantation embryos.
2021-03-15 | GSE158360 | GEO