Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptome based identification of mouse cumulus cell markers that predict the developmental competence of their enclosed antral oocytes

ABSTRACT: The cumulus cells (CCs) that envelope antral and ovulated oocytes have been regarded as putative source of non-invasive markers of the oocyte developmental competence and a number of studies have observed a correlation between CCs gene expression, embryo quality and final pregnancy outcome. With the aim of identifying marker transcripts, we isolated CCs from antral mouse oocytes of known developmental incompetence (NSN-CCs) or competence (SN-CCs) and compared their transcriptomes. Total RNA was isolated from cumulus cells derived from oocytes with a SN or NSN chromatin organization.

ORGANISM(S): Mus musculus

SUBMITTER: Alessandro Prigione

PROVIDER: E-GEOD-46906 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcriptome based identification of mouse cumulus cell markers that predict the developmental competence of their enclosed antral oocytes.

Vigone Giulia G Merico Valeria V Prigione Alessandro A Mulas Francesca F Sacchi Lucia L Gabetta Matteo M Bellazzi Riccardo R Redi Carlo Alberto CA Mazzini Giuliano G Adjaye James J Garagna Silvia S Zuccotti Maurizio M

BMC genomics 20130607

<h4>Background</h4>The cumulus cells (CCs) enveloping antral and ovulated oocytes have been regarded as putative source of non-invasive markers of the oocyte developmental competence. A number of studies have indeed observed a correlation between CCs gene expression, embryo quality, and final pregnancy outcome. Here, we isolated CCs from antral mouse oocytes of known developmental incompetence (NSN-CCs) or competence (SN-CCs) and compared their transcriptomes with the aim of identifying distinct ...[more]

PMID: 23758669

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Project description:Mammalian oogenesis is a complex mechanism entailing the maturation of primordial follicles into preovulatory follicles followed by the release of antral oocytes. In the ovary, the antral compartment consists of two populations of oocytes whose main difference regards the ability to resume meiosis and to develop, after the egg-sperm fusion, until the blastocyst stage. Still inexplicably, the antral oocytes distinguishable as Surrounded Nucleolus (SN; 70% of the antral oocytes population) are able to develop to the blastocyst stage, while those showing a Not Surrounded Nucleolus (NSN) arrest at the two-cell stage. RNA labeling and hybridization on microarray was done independently for SN and NSN oocytes with 2 replications. Two sets of 50 antral oocytes were collected in M2 medium and then were labelled with Cy3-CTP. Fluorescently labelled microarray targets were prepared using a Low RNA Input Fluorescent Linear Amplification Kit (Agilent) with 2-round amplification. Cy5-CTP-labeled reference target was produced from mixture of Stratagene Universal Mouse Reference RNA and MC1 cell RNA. Targets were purified using an RNeasy Mini Kit (Qiagen), and then quantified on a NanoDrop scanning spectrophotometer (NanoDrop Technologies). Target cRNA was hybridized to the NIA Mouse 44K Microarray v3.0 (whole genome 60-mer oligo arrays, Agilent Technology, design ID 015087) according to manufacturers protocol (Two-Color Microarray-Based Gene Expression Analysis Protocol, Product # G4140-90050, Version 5.0.1). Slides were scanned with an Agilent DNA Microarray Scanner (model G2505-64120) at 100% and 10% PMT in both channels (except for the two oocyte Samples which were only scanned once each), with a scan resolution of 5um. All hybridizations compared one Cy3-CTP-labeled experimental target to the single Cy5-CTP-labeled reference target which was used for normalization. In the final analysis, we made a cutoff of 2.0 for the processed data (i.e., if value<2.0, it is set as 2.0) to minimize the noises from 2-round linear amplifications.

Dataset Information

Transcriptome based identification of mouse cumulus cell markers that predict the developmental competence of their enclosed antral oocytes

Publications

Transcriptome based identification of mouse cumulus cell markers that predict the developmental competence of their enclosed antral oocytes.

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