Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of immature germinal vesicle stage oocytes with mature metaphase II oocytes to identify and characterize the changed and stable transcripts during mouse oocyte maturation.

ABSTRACT: 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)

ORGANISM(S): Mus musculus

SUBMITTER: You-Qiang Su

PROVIDER: E-GEOD-5668 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Selective degradation of transcripts during meiotic maturation of mouse oocytes.

Su You-Qiang YQ Sugiura Koji K Woo Yong Y Wigglesworth Karen K Kamdar Sonya S Affourtit Jason J Eppig John J JJ

Developmental biology 20060912 1

There is massive destruction of transcripts during the 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 a 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 transcrip ...[more]

PMID: 17022963

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Project description:Developmental competences of oocytes derived from prepubertal heifers are lower than those derived from adult counterparts. The objective of this study was to identify a range of genes associated with reduced oocyte competence that are differentially expressed between adult versus prepubertal donors. Microarray experiments were conducted using total RNA isolated from GV and MII stages oocytes collected from adult and prepubertal animals using Affymetrix GeneChip Bovine Genome Array containing 24,072 probe sets representing over 23,000 transcripts. A total of 549 and 333 genes were differentially expressed between prepubertal versus adult bovine MII and GV stages oocytes respectively. Out of these, 312 and 176 genes were up-regulated, while 237 and 157 were down-regulated in prepubertal when compared with adult MII and GV oocytes respectively. Ontological classification of the differentially expressed genes revealed that up-regulated genes in adult oocytes were involved in signal transduction, regulation of transcription DNA-dependent, and transport. Results from the present study indicated that significant number of genes were differentially expressed (>2-fold, p<0.01) between the two groups. Thus the decreased developmental competence of oocytes from prepubertal heifers may be induced due to difference in gene expression abundance as observed in our study. In conclusion, transcript abundance analyses of oocytes using microarray approach have been carried out in bovine and several other species. However, to our knowledge, this is the first study carried out to examine genes expression differential abundance in oocytes derived from perpubertal versus adult Japanese Black Cattle. Bovine 4b PP biological rep1, Bovine 78b PP biological rep2, Bovine 79 PP biological rep3 represents GV stage oocytes derived from Prepubertal (PP) heifer group, while Bovine 74b A biological rep1, Bovine 80b A biological rep2, Bovine 81 A biological rep3 represents GV stage oocytes derived from Adult (A) cow group. Bovine 7 PP biological rep1, Bovine 53 PP biological rep2, Bovine 57 PP biological rep3 represents MII stage oocytes derived from Prepubertal heifer group, while Bovine 59 A biological rep1, Bovine 70 A biological rep2, Bovine 71 A biological rep3 represents MII stage oocytes from Adult cow group. The ovaries of adult cows (Japanese black cattle) were collected from local abattoir while ovaries of prepubertal Japanese black heifers (9 to 12 months old) were collected by spay device at several commercial farms. The collected ovaries of both the adult cows and prepubertal heifers groups were transported to the research laboratory in 0.67% (w/v) NaCl solution containing 100 mg/L kanamycin sulfate (Meiji Seika, Tokyo, Japan). For both groups, cumulus oocyte complexes (COCs) from ovarian follicles 2 to 8M-BM- mm in diameter were aspirated by using an 18 gauge needle (Terumo co, Tokyo, Japan) attached to a 5 ml disposable syringe (Nipro, Osaka, Japan). After collection, the COCs were washed twice with Tyrode-lactate-pyruvate-polyvinylalcohol (Hepes-TLP-PVA) and TCM 199 (Invitrogen, Gibco, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (maturation medium). Only COCs with evenly granulated cytoplasm surrounded by multiple layers of compact cumulus cells were used in all the experiments. The COCs (70 to 80) were placed in 200 M-BM-5L drop of the maturation medium in petri dish (35x10mm, Becton Dickinson Labware, Oxnard, CA, USA) covered with paraffin liquid (Nacalai Tesque Inc, Kyoto, Japan) and cultured at 38.5M-BM-0C in a humidified atmosphere of 5% CO2 in air for 20 to 22 h for maturation.

Dataset Information

Transcription profiling of immature germinal vesicle stage oocytes with mature metaphase II oocytes to identify and characterize the changed and stable transcripts during mouse oocyte maturation.

Publications

Selective degradation of transcripts during meiotic maturation of mouse oocytes.

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