Unknown,Transcriptomics,Genomics,Proteomics

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Gene Expression Profiling of Single Embryos, Significant Effects of In Vitro Maturation, Fertilization and Culture

ABSTRACT: In vitro production (IVP) has been shown to affect embryonic gene expression and often result in large offspring syndrome (LOS) in cattle and sheep. To dissect the effects of in vitro maturation, fertilization and culture, we compared the expression profiles of single bovine blastocysts generated by: 1) in vitro maturation, fertilization and culture (IVF); 2) in vivo maturation, fertilization and in vitro culture (IVD); and 3) in vivo maturation, fertilization and development (AI). To conduct expression profiling, total RNA was isolated from individual embryos, linearly amplified and hybridized to a custom bovine cDNA microarray containing approximately 6,300 unique genes. There were 306, 367 and 200 genes differentially expressed between the AI and IVD, IVF and IVD and AI and IVF comparisons, respectively. Interestingly, 44 differentially expressed genes were identified between the AI embryos and both the IVF and IVD embryos, making these potential candidates for LOS. There were 60 genes differentially expressed between the IVF embryos and the AI and IVD embryos. The Gene Ontology category M-bM-^@M-^\RNA processingM-bM-^@M-^] was over-represented among the genes that were down-regulated in the IVF embryos, indicating an effect of in vitro oocyte maturation/fertilization on embryonic gene expression. A culture effect on the expression of genes involved in translation was also observed by the comparison of AI to IVD embryos. Reference design with dye swap. 43 individual embryos were analyzed.

ORGANISM(S): Bos taurus

SUBMITTER: Xiangzhong Yang

PROVIDER: E-GEOD-8807 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Gene expression profiling of single bovine embryos uncovers significant effects of in vitro maturation, fertilization and culture.

Smith Sadie L SL Everts Robin E RE Sung Li-Ying LY Du Fuliang F Page Raymond L RL Henderson Boyd B Rodriguez-Zas Sandra L SL Nedambale Tshimangadzo L TL Renard Jean-Paul JP Lewin Harris A HA Yang Xiangzhong X Tian X Cindy XC

Molecular reproduction and development 20090101 1

In vitro production (IVP) has been shown to affect embryonic gene expression and often result in large offspring syndrome (LOS) in cattle and sheep. To dissect the effects of in vitro maturation, fertilization and culture on bovine embryos, we compared the expression profiles of single blastocysts generated by: (1) in vitro maturation, fertilization and culture (IVF); (2) in vivo maturation, fertilization and in vitro culture (IVD); and (3) in vivo maturation, fertilization and development (AI). ...[more]

PMID: 18449896

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Project description:Cows were selected from two groups of 12 cows enrolled in a large experiment to assess the effects of ad libitum or restricted intake of moderate-energy diets during the entire dry period on pre-partum metabolism and post-partum metabolism and performance. A corn silage-based diet (26% of diet dry matter) providing 1.59 Mcal/kg during the far-off dry period (first 5 wk of an 8-wk dry period) or providing 1.61 Mcal/kg during the close-up dry period (last 3 wk of the dry period) was fed for ad libitum or restricted intake. Four multiparous Holstein cows were randomly selected from the ad libitum and restricted intake groups. A 7,872-element cDNA microarray spotted in duplicate on amino silane-coated glass slides (Everts et al. 2005, Vet. Immunol. Immunopathol. 105:235-245) was used for transcript profiling. Annotation was based on similarity searches (January, 2005) using sequential BLASTN and TBLASTX against human and mouse UniGene databases and the human genome. The 7,872-element array represents more than 6,300 unique genes. Gene Ontology (GO) terms were parsed from LocusLink to functionally annotate cDNA sequences. RNA (20 ug) from liver tissue and a reference standard derived from a mixture of tissues not including liver were used to make aminoallyl-labeled cDNA followed by incorporation of Cy3-ester and Cy5-ester (Amersham, Piscataway, NJ). Each experimental sample was co-hybridized with the reference standard, which allowed the treatment of fluorescence ratios as measurements of relative expression across all samples and time points. Probes were hybridized to the array for 2 days at 42 C. All samples were hybridized to duplicate slides for a total of 4 spots per cDNA element. A total of 106 microarrays were run to complete the experiment. Slides were scanned for both dye channels with a Scanarray 4000 (GSI-Lumonics, Billerica, MA) dual-laser confocal scanner and images were processed using GenePix 4.0 (Axon Instruments, Inc., CA).

Dataset Information

Gene Expression Profiling of Single Embryos, Significant Effects of In Vitro Maturation, Fertilization and Culture

Publications

Gene expression profiling of single bovine embryos uncovers significant effects of in vitro maturation, fertilization and culture.

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