Single cell analysis of sister blastomeres of parthenogenetic 2-cell mouse embryos respecting pair associations
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ABSTRACT:
PROVIDER: PRJNA437476 | ENA |
REPOSITORIES: ENA
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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.
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Project description: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
2017-08-06 | GSE94050 | GEO
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
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2023-10-17 | GSE233762 | GEO
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
Project description:In pluripotential reprogramming, a pluripotent state is established within somatic cells. In this study, we have generated induced pluripotent stem (iPS) cells from bi-maternal (uniparental) parthenogenetic neural stem cells (pNSCs) by transduction with four (Oct4, Klf4, Sox2, and c-Myc) or two (Oct4 and Klf4) transcription factors. The parthenogenetic iPS (piPS) cells directly reprogrammed from pNSCs were able to generate germline-competent himeras, and hierarchical clustering analysis showed that piPS cells were clustered more closer to parthenogenetic ES cells than normal female ES cells. Interestingly, piPS cells showed loss of parthenogenetic-specific imprinting patterns of donor cells. Microarray data also showed that the maternally imprinted genes, which were not expressed in pNSCs, were upregulated in piPS cells, indicating that pluripotential reprogramming lead to induce loss of imprinting as well as re-establishment of various features of pluripotent cells in parthenogenetic somatic cells. 5 samples were analyzed by microarray, each one them in duplicate. fNSC: Mouse female NSC (Neural Stem Cell) pNSC: Mouse parthenogenetic NSC (Neural Stem Cell) piPS-2F: Mouse parthenogenetic induced pluripotent cells derived from NSC overexpressing Oct4 and Klf4 pESC-B: Mouse parthenogenetic ESC (Embryonic Stem Cell) SSEA-1 sorted fESC: Mouse female ESC (Embryonic Stem Cell) OG2
2010-05-20 | E-GEOD-15561 | biostudies-arrayexpress
Project description:In pluripotential reprogramming, a pluripotent state is established within somatic cells. In this study, we have generated induced pluripotent stem (iPS) cells from bi-maternal (uniparental) parthenogenetic neural stem cells (pNSCs) by transduction with four (Oct4, Klf4, Sox2, and c-Myc) or two (Oct4 and Klf4) transcription factors. The parthenogenetic iPS (piPS) cells directly reprogrammed from pNSCs were able to generate germline-competent himeras, and hierarchical clustering analysis showed that piPS cells were clustered more closer to parthenogenetic ES cells than normal female ES cells. Interestingly, piPS cells showed loss of parthenogenetic-specific imprinting patterns of donor cells. Microarray data also showed that the maternally imprinted genes, which were not expressed in pNSCs, were upregulated in piPS cells, indicating that pluripotential reprogramming lead to induce loss of imprinting as well as re-establishment of various features of pluripotent cells in parthenogenetic somatic cells.
2010-01-23 | GSE15561 | GEO
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.
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Project description: Not available
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Project description:In Xenopus laevis, a number of studies identified vegetal factors that specify the germ line, endoderm and dorsal axis, but there are few studies demonstrating roles for animal-enriched maternal mRNAs. Therefore, we carried out a microarray analysis to identify novel maternal transcripts enriched in animal blastomeres. We sought to maximize differences between animal and vegetal samples. To that end, we dissected 8-cell embryos into animal blastomeres and vegetal blastomeres, and further dissected the vegetal blastomeres into vegetal-most halves (VP) and equatorial regions (discarded).
2013-12-21 | E-GEOD-48659 | biostudies-arrayexpress