Project description:Poly-ribosomal mRNA from pools of 75 bovine oocytes at the germinal vesicle(GV) and metaphase II(MII) stages were isolated by sucrose fractionation then compared through microarray hybridization.
Project description:Poly-ribosomal mRNA from pools of 75 bovine oocytes at the germinal vesicle(GV) and metaphase II(MII) stages were isolated by sucrose fractionation then compared through microarray hybridization. 3 biological replicates from germinal vesicle(GV) and metaphase II(MII) bovine oocytes were hybridized against each other while alternating dyes. One additional technical replicate (dye-swap of one of the previous hybridizations) was added to balance dye usage. The normalized log2 ratio (GV/MII) data is provided in the 'normalized_data.txt' file. Please note that data were scanned/extracted using a different Agilent reference file, and thus the feature extraction number for a given probe (in both normalized and raw data files) is different from that in the official Agilent-041017 platform (GPL18537). The reference file used for this study is provided as a Series supplementary file (041017_D_DNAFront_BCBottom_20120517.txt).
Project description:The bidirectional communication between bovine oocytes and CCs is vital for functioning and development of both cell types. We used microarray to identify genes which are differentially expressed between germinal vesicle (GV)- and metaphase II (MII)-stage oocytes and CCs and those differentially expressed when oocytes mature with or without the other. We also identified genes differentially expressed between CCs at GV and MII stages.
Project description:The statement that fully-grown porcine oocytes are transcriptionally quiescent is not as strong as before. Currently, we know that there is a difference in the transcription profile of germinal vesicle (GV) and metaphase II (MII) oocytes. The goal of our study was to compare the transcription profile of GV, germinal vesicle breakdown (GVBD), metaphase I (MI), and metaphase (MII) oocytes cultivated in a chemically defined, FLI medium. Oocytes were sequenced and subsequently validated using quantitative reverse transcription polymerase chain reaction (RT-qPCR).
Project description:The bidirectional communication between bovine oocytes and CCs is vital for functioning and development of both cell types. We used microarray to identify genes which are differentially expressed between germinal vesicle (GV)- and metaphase II (MII)-stage oocytes and CCs and those differentially expressed when oocytes mature with or without the other. We also identified genes differentially expressed between CCs at GV and MII stages. Slaughterhouse ovaries were collected and GV-stage cumulus oocyte complexes (COCs) were aspirated. Different stages and types of oocytes and CCs were used for total RNA isolation and hybridisation on Affymetrix microarray.
Project description:Polyadenylated RNA from individual germinal vesicle and metaphase II ooyctes was amplified and processed for Illumina sequencing. Increases in polyadenylated transcript abundance were observed and are likely due to cytoplasmic polyadenylation. Transcriptomic analysis of GV and MII bovine oocytes.
Project description:In cattle, almost all fully grown vesicle stage oocytes (GV) have the ability to resume meisos, develop to Metaphase II stage (MII), support fertilization and progress through the early embryonic cycles in vitro. Yet without intensive selection, the majority fail to develop to the blastocyst stage. Using the Affymetrix Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of the bovine oocyte and to identify the key pathways associated with oocyte meiotic maturation and developmental potential.
Project description:We aimed to reveal gene expression profiles of human oocytes at each maturation stage by single cell RNA-seq analyses. We investigated transcriptomes of immature oocytes at the germinal vesicle (GV) stage, maturating oocytes at the metaphase I (MI) stage and in vitro matured oocytes at the metaphase II (MII) stage.
Project description:In cattle, almost all fully grown vesicle stage oocytes (GV) have the ability to resume meisos, develop to Metaphase II stage (MII), support fertilization and progress through the early embryonic cycles in vitro. Yet without intensive selection, the majority fail to develop to the blastocyst stage. Using the Affymetrix Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of the bovine oocyte and to identify the key pathways associated with oocyte meiotic maturation and developmental potential. Immature and in vitro matured bovine oocytes were collected for RNA extraction and hybridization on Affymetrix GeneChip Bovine Genome array. Careful removal of cumulus and selection of oocytes was carried out under the stereo microscope in order to examine the actual cumulus-free temporal oocyte gene expression profiles. Immature oocytes at time 0 h and in vitro matured oocytes at 24 h were collected for analysis.
Project description:There is massive destruction of transcripts during maturation of mouse oocytes. The objective of this project was to identify and characterize the transcripts that are degraded versus those that are stable during the transcriptionally silent germinal vesicle (GV)-stage to metaphase II (MII)-stage transition using the microarray approach. A system for oocyte transcript amplification using both internal and 3'-poly(A) priming was utilized to minimize the impact of complex variations in transcript polyadenylation prevalent during this transition. Transcripts were identified and quantified using Affymetrix Mouse Genome 430 v2.0 GeneChip. The significantly changed and stable transcripts were analyzed using Ingenuity Pathways Analysis and GenMAPP/MAPPFinder to characterize the biological themes underlying global changes in oocyte transcripts during maturation. It was concluded that the destruction of transcripts during the GV to MII transition is a selective rather than promiscuous process in mouse oocytes. In general, transcripts involved in processes that are associated with meiotic arrest at the GV-stage and the progression of oocyte maturation, such as oxidative phosphorylation, energy production, and protein synthesis and metabolism, were dramatically degraded. In contrast, transcripts encoding participants in signaling pathways essential for maintaining the unique characteristics of the MII-arrested oocyte, such as those involved in protein kinase pathways, were the most prominent among those stables. Experiment Overall Design: Comparison immature GV-stage oocyte (3 biological replicates) with mature MII-stage oocytes (3 biological replicates)