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| Status |
Public on Jun 01, 2012 |
| Title |
Gene Expression profile in preimplantation in vitro embryos derived from Prepubertal heifers and Adult Cattle |
| Organism |
Bos taurus |
| Experiment type |
Expression profiling by array
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| Summary |
Age of the donor animal has a significant influence on the oocyte’s ability to complete maturation and acquire the mRNA and proteins required for normal embryonic development. It is well known that developmental competence is reduced in oocytes derived from juvenile animals when compared to that derived from adults. However, the molecular mechanisms associated with these differences are not well elucidated. The aim of this study was to analyze the transcriptome differences between adult versus prepubertal derived preimplantation embryos and to identify genes associated with oocyte developmental potential. We performed cDNA microarray analysis on populations of preimplantation embryos (8- to 16-cell and Day 7 blastocysts) derived from adult versus prepubertal Japanese Black cattle. Total RNA was extracted and amplified in a linear fashion and then subjected to the microarray hybridization using the Affymetrix GeneChip Bovine Genome Array. The Bovine Array contains 24 072 probe sets representing over 23000 transcripts and 19000 UniGene clusters.
Following normalization of the microarray data, analysis revealed differences in gene expression abundance in 8- to 16-cell and blastocysts harvested from adult versus prepubertal heifers. Results indicated that a total of 591 and 490 genes were differentially expressed (≥2-fold) in 8- to 16-cell and blastocyst respectively between the two groups (p<0.01). In 8- to 16-cell stage, 373 genes were up-regulated and 218 gens down-regulated in adult when compared to the prepubertal heifer group. In case of blastocyst stage embryos, 242 genes were up-regulated and 248 genes were down-regulated in adult when compared to prepubertal group.
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| Overall design |
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 ovaries were transported to the research laboratory in 0.67% (w/v) NaCl solution containing 100 mg/L kanamycin sulfate. COC were placed in 200 µL drop of the maturation medium (TCM 199 + 10% FBS) and cultured at 38.5°C in a humidified atmosphere of 5% CO2 in air for 20 to 22 h for in vitro maturation (IVM). After 20 to 22 h IVM, the COC were fertilized in vitro with frozen–thawed semen of Japanese Black bull in BO medium. Six hours after IVF, oocytes were denuded of surrounding cumulus cells by gentle pipetting in a solution of 0.1% (w/v) hyaluronidase. The presumptive zygotes were cultured in CR1aa medium supplemented with fatty acid-free BSA (Sigma) for 2 days and cultured in CR1aa medium supplemented with 10% (v/v) fetal bovine serum (FBS) for next 5 days at 38.5 °C in 5% CO2, 5% O2 and 90% N2 with high humidity (15 to 20 zygotes per 50 µl microdrop). 8- to 16-cell refers to day 3, 70 to 72 hours post insemination( hpi) preimplantation embryo stage containing 8 to 16 cells, while blastocyst refer to pre-hatching day 7 expanded blastocysts. Pools of 8- to 16- cell embryos (50 in each sample) and blastocysts (10 in each sample) from prepubertal and adult groups were used for cDNA microarray.
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| Contributor(s) |
Dorji D, MIYOSHI K, YOSHIDA M |
| Citation missing |
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| Submission date |
Oct 08, 2010 |
| Last update date |
Dec 17, 2012 |
| Contact name |
Dorji Dorji |
| E-mail(s) |
dr.dorji@gmail.com
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| Phone |
+818039465965
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| Organization name |
Kagoshima University
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| Department |
Faculty of Agriculture
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| Lab |
Laboratory of Animal Reproduction
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| Street address |
Korimoto
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| City |
Kagoshima |
| ZIP/Postal code |
890-0065 |
| Country |
Japan |
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| Platforms (1) |
| GPL2112 |
[Bovine] Affymetrix Bovine Genome Array |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA132383 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE24596_RAW.tar |
17.9 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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