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Single embryo RNA-seq of control and hydroxyurea treated embryos produced by nuclear transfer of cumulus cells. Single cell RNAseq of cumulus cells was performed using the same protocol.

ABSTRACT: Single embryo RNA-seq of control and hydroxyurea treated embryos produced by nuclear transfer of cumulus cells. Single cell RNAseq of cumulus cells was performed using the same protocol.

PROVIDER: PRJNA700563 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13649918_1.fastq.gz	Fastqsanger.gz
	SRR13649918_2.fastq.gz	Fastqsanger.gz
	SRR13649919_1.fastq.gz	Fastqsanger.gz
	SRR13649919_2.fastq.gz	Fastqsanger.gz
	SRR13649920_1.fastq.gz	Fastqsanger.gz

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Single embryo RNA-seq of control and hydroxyurea treated embryos produced by nuclear transfer of cumulus cells. Single cell RNAseq of cumulus cells was performed using the same protocol.

Project description:Slow fork speed inducer, hydroxyurea, efficiently improved development of somatic cell nuclear transfered embryos to the blastocyst stage. Here, we performed single embryo RNA-seq approach to investigate whether nuclear transfer embryos exhibited a transcriptional programme similar to in vivo fertilised embryos.

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The bovine cumulus proteome of cumulus oocyte complexes that successfully matured or failed to mature after maturation in-vitro and in-vivo

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Analysis of the proteome of individual equine cumulus complexes after maturation in-vitro and in-vivo

Project description:Maturation of oocytes under in-vitro conditions (IVM) results in impaired developmental competence compared to oocytes matured in-vivo. Oocytes are closely coupled to their cumulus complex (COC) with a bidirectional exchange of metabolites. Therefore, elucidation of aberrations in cumulus metabolism in-vitro is crucial for a better mimicking of physiological maturation conditions. The aim of this study was the analysis of the equine cumulus cells proteome of single cumulus complexes of metaphase II oocytes matured either under in-vivo (n=8) or in-vitro (n=7) conditions. For in-vivo COC collection mares were slaughtered 30 hours after injection and cumulus complexes from the dominant follicle were harvested for analysis. For in-vitro maturation COCs were recovered from mares out of oestrous and matured for 30 hours in-vitro. COCs were separated in cumulus complexes and oocytes, and only cumulus of successfully matured oocytes was analyzed for this study. All cumulus samples were washed, snap frozen and stored in liquid nitrogen until preparation for analysis.

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Bovine adult versus prepubertal cumulus cells

Project description:The present study was undertaken to discover molecular markers in bovine cumulus cells predictive of oocyte competence and elucidate their functional significance. Differences in RNA transcript abundance in cumulus cells harvested from oocytes of adult versus prepubertal animals (model of poor oocyte quality) were identified by microarray analysis. Four genes of interest encoding for the lysosomal cysteine proteinases cathepsin B, S, K and Z and displaying greater transcript abundance in cumulus cells surrounding oocytes harvested from prepubertal animals were chosen for further investigation. Greater mRNA abundance for such genes in cumulus cells of prepubertal oocytes was confirmed by real time RT-PCR. Elevated transcript abundance for cathepsins B, S and Z was also observed in cumulus cells surrounding adult metaphase II oocytes that developed to the blastocyst stage at a low percentage following parthenogenetic activation, versus those that developed at a high percentage. Functional significance of cumulus cell cathepsin expression to oocyte competence was confirmed by treatment of cumulus oocyte complexes during in vitro oocyte maturation with a cell permeable cysteine proteinase (cathepsin) inhibitor. Inhibitor treatment decreased apoptotic nuclei in the cumulus layer and enhanced development of parthenogenetically activated and in vitro fertilized adult oocytes to the blastocyst stage. Stimulatory effects of inhibitor treatment during meiotic maturation on subsequent embryonic development were not observed when oocytes were matured in the absence of cumulus cells. Results support a functional role for cumulus cell cathepsins in compromised oocyte competence and suggest that cumulus cell cathepsin mRNA abundance may be predictive of oocyte quality. Keywords: Bovine, microarray, cDNA, cumulus cells

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Project description:The maturation of an oocyte, the female gamete, during folliculogenesis is highly dependent on the molecular interaction with the somatic cells. Here we generated transcriptome data of single oocytes and the surrounding cumulus cells from antral follicles, and developed an analytical framework to unveil their inter-cellular communication. We identified hundreds of ligand-receptors pairs that can transduce paracrine signaling between oocyte and cumulus. Several of the ligand-coding genes expressed in oocytes and cumulus cells are also functionally associated with regulation of transcription. Ligand-coding genes expressed in oocytes or cumulus showed distinct enrichment for biological functions that are likely associated with a coordinated formation of the transzonal projections from the cumulus that reach the oocyte’s membrane. Finally, there were thousands of gene pairs with non-trivial linear co-expression pattern between oocytes and cumulus cells. Our analyses revealed a complex and functional regulatory circuit between the oocyte and surrounding cumulus cells.

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Multicenter Study of Hydroxyurea (MSH)

Project description:Multicenter Study of Hydroxyurea (MSH)

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Multicenter Study of Hydroxyurea (MSH)

Project description:Multicenter Study of Hydroxyurea (MSH)

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