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Single cell analysis of sister blastomeres of 2-cell mouse embryos respecting pair associations

ABSTRACT: Upon 2-cell embryo splitting, individual blastomeres were compared and contrasted with each other respecting pair associations (e.g. blastomere '1a' and '1b' of embryo 1, '2a' and '2b' of embryo 2, and so forth) Transcriptome analysis followed by cluster analysis (Ward) was able to match a minority of the blastomeres with the correct sister blastomere

ORGANISM(S): Mus musculus

PROVIDER: GSE94050 | GEO | 2017/08/06

SECONDARY ACCESSION(S): PRJNA368650

REPOSITORIES: GEO

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Single embryo analysis of monozygotic twin blastocysts of the mouse respecting pair associations

Project description:Upon 2-cell embryo splitting, individual blastomeres were cultured in KSOM(aa) medium and developed into blastocysts, which were compared and contrasted with each other respecting pair associations (e.g. twin vs cotwin) Transcriptome analysis revelaled minimal differences between twins and unmanipulated controls, in contrast to larger differences between twins and cotwins; these differences are mainly related to the epiblast lineage

2017-08-06 | GSE90674 | GEO

Single cell analysis of sister blastomeres of parthenogenetic 2-cell mouse embryos respecting pair associations

Project description:Following fertilization in mammals, it is generally accepted that totipotent cells are exclusive to the zygote and to each of the two blastomeres originating from the first mitotic division. We counter that this classic view needs to be revised, because we have presented compelling evidence that the sister blastomeres are both totipotent in only a subset of 2-cell stage mouse embryos (PMID 28811525). Building on our previous findings, we here ask the question if the interblastomere differences depend - at least in part - on the contribution of sperm, since the area of sperm entry is inherited preferentially by one blastomere. To this end, we created sister 2-cell stage blastomeres without sperm entry point, by parthenogenesis. We compare the transcriptomes of the sister blastomeres, to see if the interblastomere mRNA differences observed after fertilization (GSE94050) are still there when the sperm entry point is missing.

2019-07-10 | GSE111589 | GEO

Single cell analysis of sister blastomeres of parthenogenetic 2-cell mouse embryos respecting pair associations

Project description:Single cell analysis of sister blastomeres of parthenogenetic 2-cell mouse embryos respecting pair associations

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Transcript profiling of individual twin blastomeres derived by splitting 2-cell stage murine embryos

Project description:We have examined whether twin blastomeres from 2-cell stage mouse embryos differ in mRNA content. Amplified mRNA from 12 blastomeres derived from six embryos approximately mid-way through their second cell cycle was analyzed. Probes displaying normalized values greater than 0.25 were selected and examined for consistent bias in expression within blastomere pairs. Although transcript content varied both between individual embryos and twin blastomeres, no consistent asymmetries were observed for the majority of genes. On the other hand, 769 genes displayed a greater than 1.4-fold difference in expression across 5 of 6 pairs of blastomeres and 178 genes differed across all 6 pairs. These genes separated into two groups by class discovery clustering. Of the 769 differentially expressed genes, 163 were significantly up- or down-regulated in one sister blastomere compared to the other. Transcripts encoding proteins implicated in RNA processing and cytoskeletal organization were highly represented among the most abundant, differentially distributed mRNA. We conclude that there are many differences that distinguish twin blastomeres derived from a single 2-cell stage embryo but that only a few of these differences are consistent across multiple pairs of embryos. We hypothesize that a stochastically-based lack of synchrony in cell cycle progression between the two cells might explain some or all of the asymmetries in transcript composition. 6 pairs of blastomeres were analyzed for a total of 12 samples.

2010-05-28 | E-GEOD-21688 | biostudies-arrayexpress

Transcript profiling of individual twin blastomeres derived by splitting 2-cell stage murine embryos

2010-05-15 | GSE21688 | GEO

Totipotency of mouse zygotes extends to single blastomeres of 4-cell stage embryos

Project description:In multicellular organisms, heterogametes of oocytes and sperms are fertilized and resulting zygotes give rise to new individuals. The ability of zygotes that produce a fully formed individual from single cell when placed in a supportive environment is defined as totipotency. Given that totipotent cells are the source of all multicellular organisms, better understanding of totipotency has a profound effect on not only biology but also our society. However, the exact distribution of totipotent cells in mammals remains elusive, although zygotes and single blastomeres at 2-cell stage embryos has been thought to be only mouse cells to be totipotent. We now show that a single blastomere isolated from 2- and 4-cell stage embryos gives rise to a fertile adult individual when it placed in a uterus, although isolation of blastomeres at these stage results in the disturbance of transcriptome in single blastomere derived embryos. Single blastomeres from 8-cell and morula stage embryos that were separately cultured in vitro exhibited severe defects in the formation of epiblast and primitive endoderm in the inner cell mass and the development to blastocysts, respectively. Our results thus indicate that totipotency of mouse zygotes extends to single blastomeres of 4-cell stage embryos.

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Defining Cell Fate And Embryonic Genome Activation By Global Single-Cell cDNA Analysis of Blastomeres From 5-to 8-Cell Human Embryos.

2010-10-30 | GSE22032 | GEO

Defining Cell Fate And Embryonic Genome Activation By Global Single-Cell cDNA Analysis of Blastomeres From 5-to 8-Cell Human Embryos.

Project description:To determine blastomere fate and embryonic genome activation (EGA) at 5- to 8-cell stage human embryos by global gene expression profile of amplified cDNA from blastomeres at the single cell level Forty-nine blastomeres from 5-, 6- and 8-cell human embryos were analyzed through whole genome wide analysis following an efficient cDNA amplification protocol (Kurimoto et al., 2007) with slight modifications. Single biopsied blastomeres were also compared with two amplified inner cell masses and two trophectoderms from blastocysts.

2010-10-30 | E-GEOD-22032 | biostudies-arrayexpress

Filter aided, single tip (FAST) method for high throughput, ultrasensitive proteomics sample preparation reveals proteome asymmetry in Xenopus embryos

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2022-07-12 | PXD015175 | Pride

Cell fate inclination within 2-cell and 4-cell mouse embryos revealed by single-cell RNA sequencing

Project description:It remains an open question when and how the first cell fate decision is made in mammals. Using deep single-cell RNA-seq of matched sister blastomeres, we report highly reproducible interblastomere differences among ten 2-cell and five 4-cell mouse embryos. Inter-blastomere gene expression differences dominated between-embryo differences and noises, and were sufficient to cluster sister blastomeres into distinct groups. Dozens of protein-coding genes exhibited reproducible bimodal expression in sister blastomeres (0 vs. 1e3-1e6 of FPKM), which cannot be explained by random fluctuations. The protein expression of one of these bimodal genes, Gadd45a, exhibited clear inter-blastomeric contrasts. We traced some of the bimodal mRNA expressions to embryonic genome activation, and others to blastomere-specific RNA depletion. Inter-blastomere differences created co-expression gene networks that were much stronger and larger than those that can be possibly created by random noises. The highly correlated gene pairs at the 4-cell overlapped with those showing the same directions of differential expression between inner cell mass (ICM) and trophectoderm (TE). These data substantiate the hypothesis of inter-blastomere differences in 2- and 4-cell mouse embryos, and associate these differences with ICM/TE differences.

2014-07-29 | GSE57249 | GEO

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