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Functional study of Rbbp5 in preimplantation embryos

ABSTRACT: Functional study of Rbbp5 in preimplantation embryos

PROVIDER: PRJNA763493 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15903800_1.fastq.gz	Fastqsanger.gz
	SRR15903800_2.fastq.gz	Fastqsanger.gz
	SRR15903801_1.fastq.gz	Fastqsanger.gz
	SRR15903801_2.fastq.gz	Fastqsanger.gz
	SRR15903802_1.fastq.gz	Fastqsanger.gz

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Functional study of Rbbp5 in preimplantation embryos

Project description:We performed RNA-sequencing, ChIP-sequncing and Brif-sequencing on Rbbp5 knockout (KO) and wild-type (WT) mouse ICM and E5.0 epiblast. Our results showed that Rbbp5 KO lead to significant reduction of H3K4me3 modification at early blastocyst stage. The change in H3K4me3 landscape occured before the transcriptome change. Moreover, Rbbp5 KO specifically affected epiblast lineages the most at late blastocyst stage and the KO embryos died at E5.5.

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Gene Expression Human Preimplantation Embryos

Project description:Differential gene expression in preimplantation embryos has been documented, but few focused studies have been done to compare differential expression in human embryos after embryonic genome activation and specifically how they relate to blastocyst development. We hypothesized that blastocyst stage embryos would differentially express genes in pathways important in cell division, mobilization, and processes important in embryo implantation including endometrial apposition, adhesion, and invasion. We analyzed gene expression in 6 preimplantation human embryos. Embryos studied were previously cryopreserved, supernumerary human embryos donated by couples who completed their family building through in vitro fertilization and had given specific consent for use in research. Embryos cryopreserved at the pronuclear stage were thawed and cultured to cleavage (Day 3) or blastocyst (Day 5) stage. Differential gene expression was first obtained through Affymetrix gene expression microarrays and then validated both in silico using the Gene Expression Omnibus and in vitro with RT-qPCR. Compared to cleavage stage embryos, blastocyst stage embryos differentially expressed 51 genes (p < 0.001), with overrepresentation in amoebiasis pathways and pathways in cancer.

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Project description:Upon fertilization, extensive chromatin reprogramming occurs during preimplantation development. Growing evidence reveals species-dependent regulations of this process in mammals. ATP-dependent chromatin remodeling factor SMARCA5 (also known as SNF2H) is required for peri-implantation development in mice. However, the specific functional role of SMARCA5 in preimplantation development and if it is conserved among species remain unclear. Herein, comparative analysis of public RNA-seq datasets reveals that SMARCA5 is universally expressed during oocyte maturation and preimplantation development in mice, cattle, humans and pigs with species-specific patterns. Immunostaining analysis further describes the temporal and spatial changes of SMARCA5 in both mouse and bovine models. siRNA-mediated SMARCA5 depletion reduces the developmental capability and compromises the specification and differentiation of inner cell mass in mouse preimplantation embryos. Indeed, OCT4 is not restricted into the inner cell mass and the formation of epiblast and primitive endoderm disturbed with reduced NANOG and SOX17 in SMARCA5-deficient blastocysts. RNA-seq analysis shows SMARCA5 depletion causes limited effects on the transcriptomics at the morula stage, however, dysregulates 402 genes, including genes involved in transcription regulation and cell proliferation at the blastocyst stage in mice. By comparison, SMARCA5 depletion does not affect the development through the blastocyst stage but significantly compromises the blastocyst quality in cattle. Primitive endoderm formation is greatly disrupted with reduced GATA6 in bovine blastocysts. Overall, our studies demonstrate the importance of SMARCA5 in fostering the preimplantation development in mice and cattle while there are species-specific effects.

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