Project description:This SuperSeries is composed of the following subset Series: GSE23033: Polycomb function during oogenesis is required for mouse early embryonic development (germinal vesicle oocytes) GSE28710: Polycomb function during oogenesis is required for mouse early embryonic development (2-cell embryos) Refer to individual Series
Project description:Comprehensive quantitative proteomic study of human pre-implantation embryo stages reveal dynamic proteome landscape from M2, 8-cell and blastocyst stage, and during trophoblast stem cell (TS) differentiation. Identified key factors in early human embryos and lineage-specific trophoblast proteome profiles, correlated with transcriptomic analyses. This direct proteomic analysis provides a comprehensive analysis of the dynamic protein expression in human embryos during pre-implantation development and a powerful resource to enable further mechanistic studies on human trophoblast development and function.
Project description:The piRNA pathway controls transposon expression in animal germ cells, thereby ensuring genome stability over generations. piRNAs are maternally deposited and required for proper transposon silencing in adult offspring. However, a long-standing question in the field is the precise function of maternally deposited piRNAs and its associated factors during embryogenesis. Here, we probe the spatio-temporal expression patterns of several piRNA pathway components during early stages of development. Amongst those, factors required for transcriptional gene silencing (TGS) showed ubiquitous abundance in somatic and pole cells throughout the first half of embryogenesis. We further analysed the transcriptomes of various embryo stages and correlated these with the presence of selected chromatin marks. We found that a number of transposon families show bursts of transcription during early embryonic stages. Transposons heavily targeted by maternally deposited piRNAs accumulated repressive chromatin marks following their spike in expression. Furthermore, depletion of maternally deposited Piwi protein in early embryos resulted in increased expression of transposons targeted by inherited piRNAs and was accompanied by a strong loss of repressive chromatin marks at coding sequences. Overall, our data suggests a pivotal role for the piRNA pathway in transposon defence during Drosophila embryogenesis in somatic cells.
Project description:During oogenesis, RNAs and proteins from maternal genome will be accumulated and they are the most important regulatory factors in early embryonic development. Parthenogenesis provides a unique source to investigate the influence of maternal genomes in early mammalian development and is proposed as an experimental tool to investigate embryo development which may solve many of the ethical concerns. Using label-free quantitative mass sepctrometry (MS), we systematically monitored protein expression profiles from six stages during pre-implantation development used by parthenogenesis model of mouse: pronucleus-, 2-cell, 4-cell, and 8-cell embryo, morula, and blastocyst. They are labeled as PA, PA-2, PA-4, PA-8, PAMO, PABL, respectively. For each stage, 6,000 embryos were used, and the experiment was performed in three biological replicates, and they had 2,048 proteins in common.
Project description:Preimplantation development is a crucial step for successful implantation and pregnancy. Although both compaction and blastocyst formation have been extensively studied, mechanisms regulating early cell division stages before compaction have remained unclear. Here, we show that ERK MAP kinase function is required for early embryonic cell division and normal cell-cell adhesion before compaction. Our analysis demonstrates that inhibition of ERK activation in the late 2-cell stage embryos leads to a reversible arrest in G2 phase in the 4-cell stage. The G2 arrested, 4-cell stage embryos show weakened cell-cell adhesion as compared to control embryos. Remarkably, microarray analyses show that most of the programmed changes of upregulated and downregulated gene expression during the 4- to 8-cell stages normally proceed in the 4-cell stage-arrested embryos, except for a portion of the genes whose expression profiles closely parallel the stages of embryonic development when arrested in G2 and released to resume development. These parallel genes include the genes encoding intercellular adhesion molecules, whose expression is found to be positively regulated by the ERK pathway. We also show that while ERK inactivation in the 8-cell stage embryos does not lead to cell division arrest, it does cause cell division arrest when cadherin-mediated cell-cell adhesion is disrupted. These results demonstrate an essential role of ERK function in the G2/M transition and the expression of adhesion molecules during the 2-cell to 8-cell stage embryos, and suggest a loose parallelism between the gene expression programs and the developmental stages before compaction. Experiment Overall Design: We examined expression profiles of genes during early cell division stages in mouse preimplantation development. We performed the genome-wide analysis by using Affymetrix GeneChip oligonucleotide microarrays, and examined the effect of the ERK pathway inhibitor U0126 on the expression profiles. Two independent experiments were carried out. We collected embryos at six points as follows; control embryos at day 1.5 (cont. 1.5, 2-cell stage), day 2.5 (cont. 2.5, 4- to 8-cell), and day 3.5 (cont. 3.5, morula to blastocyst), and the U0126-treated embryos at day 2.5 (U2.5, 4-cell arrested), embryos released from the U0126-induced arrest at day 3.5 (U3.5, 8-cell) and at day 4.5 (U4.5, morula to blastocyst). Hybridization was carried out with the Mouse Genome 430 2.0 array following Affymetrix instructions. Hybridized arrays were scanned using an Affymetrix GeneChip Scanner. Expression analysis was performed using GeneChip Operating Software v. 1.2 (GCOS) and GeneSpring 7.3.
Project description:In mouse development, long-term silencing by CpG island DNA methylation is specifically targeted to germline genes, however the molecular mechanisms of this specificity remain unclear. Here we demonstrate that the transcription factor E2F6, a member of the polycomb repressive complex 1.6 (PRC1.6), is critical to target and initiate epigenetic silencing at germline genes in early embryogenesis. Genome-wide, E2F6 binds preferentially to CpG islands in embryonic cells. E2F6 cooperates with MGA to silence a subgroup of germline genes in mouse embryonic stem cells and in vivo, a function that critically depends on the E2F6 marked box domain. Inactivation of E2f6 leads to a failure to deposit CpG island DNA methylation at these genes during implantation. Furthermore, E2F6 is required to initiate epigenetic silencing in early embryonic cells but becomes dispensable for the maintenance in differentiated cells. Our findings elucidate the mechanisms of epigenetic targeting of germline genes and provide a paradigm for how transient repression signals by DNA-binding factors in early embryonic cells are translated into long term epigenetic silencing during mammalian development.