Single-cell RNA-seq of blastomeres from 2- to 32-cell stage mouse embryos
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB12525 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR1229130_1.fastq.gz | Fastqsanger.gz | |||
| ERR1229130_2.fastq.gz | Fastqsanger.gz | |||
| ERR1229140_1.fastq.gz | Fastqsanger.gz | |||
| ERR1229140_2.fastq.gz | Fastqsanger.gz | |||
| ERR1229150_1.fastq.gz | Fastqsanger.gz |
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Project description:Transcriptomes were determined for all blastomeres of 28 embryos at the 2- (n=8), 4- (n=16) and 8-cell (n=4) stages, and for individual cells taken from 16- (n=6) and 32- (n=6) cell stage embryos. We also carefully monitored the 2- to 4-cell divisions noting whether the cleavage plane was meridional (M, along the Animal-Vegetal (AV) axis marked by the second attached polar body) or equatorial (E, bisecting the AV axis) and the order in which such divisions occurred. This resulted in four groups of 4-cell stage embryos: ME, EM, MM and EE, which were all collected 10 hours after the first 2- to 4-cell division.
2016-03-23 | E-MTAB-3321 | biostudies-arrayexpress
Project description:Nuclear pore complex (NPC) structure and maturity changed during early development before MBT in zebrafish embryos. Aiming to further confirm the difference of NPCs among different stages during MZT, we first isolated nuclei from 32-cell and 512-cell stage embryos as most zygotic genes have not been transcribed at these two stages yet. The lysates from identical number (about 12,500 nuclei per stage) isolated nuclei were then trypsin-digested and fractionated, followed by high-resolution LC-MS/MS analysis on the Q-Exactive HF-X mass spectrometers using a label-free assay. Whole blastomeres from 32-cell and 512-cell were also prepared with yolk removal to serve as total protein profile control, and the same amount lysates were analyzed by high-resolution LC-MS/MS with TMT labelled. Each sample for MS analysis had two repeats. Correlation analysis revealed that the abundance ratio from isolated Nuclei and whole deyolking balstomeres did not correlate well, but samples in Nuclei or Whole blastomeres were well correlated. This result indicates the convincible sample preparation as well as the variability in nuclei composition in early developmental period before MBT.
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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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Project description:Totipotency of mouse zygotes extends to single blastomeres of 4-cell stage embryos
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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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