Single-cell RNA-seq Analyses of Oocytes and Zygotes Reveal Earlier Maternal to Zygote Transition in Mice and Rats
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1214818 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR32095700_1.fastq.gz | Fastqsanger.gz | |||
| SRR32095700_2.fastq.gz | Fastqsanger.gz | |||
| SRR32095701_1.fastq.gz | Fastqsanger.gz | |||
| SRR32095701_2.fastq.gz | Fastqsanger.gz | |||
| SRR32095702_1.fastq.gz | Fastqsanger.gz |
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Project description:The maternal to zygote transition in mammals has been an area of intensive research over the past few decades, with an ever-changing landscape of understanding that has accompanied the rapid development of cell-profiling technology. Utilizing a full-length single cell RNA-seq protocol, we profiled mature oocytes and zygotes of mice and rats to uncover elusive transcriptomic dynamics during the maternal to zygote transition. We confirm the existence of early gene expression in the mouse zygote while revealing a similar chain of events occurring in the rat zygote. We observe an increase in nascent transcription in both species. Moreover, we find subtle but pervasive signals of differential splicing of genes related to key early zygotic activities occurring in both species. Meanwhile, we find distinct profiles of alternative polyadenylation between zygotes and oocytes in both species, particularly, in genes related to major processes within the zygotes. Finally, although a more dynamic transcriptomic landscape exists in the mouse zygote, the rat zygote also displays similar transcriptomic features, suggesting that minor zygotic activation in rat occurs earlier than originally thought.
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Project description:PABPN1L participates in the process of post-transcriptional regulation and degradation of maternal mRNA.So,we collect samples of each period from pabpn1l ko and wt female mice during different stages.Then, RNA was extracted and detected.Reverse transcription was performed on the samples that met the effective concentration.After quality inspection, build the library.GV oocytes, MII oocytes and zygotes were collected from three different PABPN1L KO or WT mice separately.
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Project description:The epigenomes of mammalian sperm and oocytes, characterized by gamete-specific 5-methylcytosine (5mC) patterns, are reprogrammed in early embryogenesis to establish full developmental potential. It is broadly accepted that the paternal genome is actively demethylated in the zygote while the maternal genome undergoes passive demethylation thanks to DNA replication over the subsequent cleavage divisions. Here we reveal that both maternal and paternal genomes undergo widespread active and passive demethylation in the pronuclear zygote before the first mitotic division. Whereas the passive demethylation requires DNA replication, the active demethylation relies on enzymatic oxidation of 5mC, as deletion of the DNA dioxygenase, Tet3, but not the inhibition of replication, blocks the active demethylation. At actively demethylated loci, 5mCs appear to be processed to unmodified cytosines in a manner independent of the DNA glycosylase TDG. These observations suggest the occurrence of genuine active demethylation in both parental genomes following fertilization. An extra supportive Tet3 knock-out female pronuclear sample related to experiment Series GSE56650.
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