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| Status |
Public on Jun 05, 2007 |
| Title |
Microarray analysis of NL and PCOS oocytes |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Polycystic ovary syndrome (PCOS), the most common cause of anovulatory infertility, is characterized by increased ovarian androgen production, arrested follicle development, and is frequently associated with insulin resistance. These PCOS phenotypes are associated with exaggerated ovarian responsiveness to FSH and increased pregnancy loss. To examine whether the perturbations in follicle growth and the intrafollicular environment affects development of the mature PCOS oocyte, genes that are differentially expressed in PCOS compared to normal oocytes were defined using microarray analysis. This analysis detected approximately 8000 transcripts. Hierarchical clustering and principal component analysis revealed differences in global gene expression profiles between normal and PCOS oocytes. 374 genes had a statistically-significant increase or decrease in mRNA abundance in PCOS oocytes. A subset of these genes was associated with chromosome alignment and segregation during mitosis and/or meiosis, suggesting that increased mRNAs for these proteins may negatively affect oocyte maturation and/or early embryonic development. Of the 374 differentially expressed genes, 68 contained putative androgen receptor, retinoic acid receptor, and/or peroxisome proliferating receptor gamma binding sites, including 9 of the genes involved in chromosome alignment and segregation. These analyses demonstrated that normal and PCOS oocytes that are morphologically indistinguishable and of high quality exhibit different gene expression profiles. Furthermore, altered mRNA levels in the PCOS oocyte may contribute to defects in meiosis and/or mitosis which might impair oocyte competence for early development and therefore contribute to poor pregnancy outcome in PCOS.
Keywords: disease state analysis
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| Overall design |
A single MII oocyte, defined by one polar body in the perivitelline space and no visible nuclear structure in the cytoplasm, was collected from 6 individual NL and 6 individual PCOS ovaries, placed immediately in TRIzol (Sigma, St. Louis MO), and stored at -80 C until further study. Total RNA was isolated from each oocyte and subjected to three rounds of linear amplification with the Ovation Biotin RNA Amplification and Labeling System (NuGen Technologies, San Carlos CA) per the manufacturer’s instructions. RNA from the GeneChip Eukaryotic Poly-A RNA Control Kit (Affymetrix, Santa Clara CA) was amplified and labeled under the same conditions for a positive control. Affymetrix GeneChip Human Genome U133 Plus 2.0 microarray chips (Affymetrix, Santa Clara, CA) were hybridized at the University of Pennsylvania Microarray Core Facility. Briefly, the linear-amplified, biotin-labled cDNA from 6 NL (N1-N6) and 6 PCOS (P1-P6) oocytes was hybridized to individual Affymetrix U133 chips. The fluorescence intensity of each chip was normalized to a trimmed mean signal of 150. Each transcript on the U133 chip was defined as present or absent in each oocyte sample using the Affymetrix Microarray Suite 5.0.
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| Contributor(s) |
Wood JR, Dumesic DA, Abbott DH, Strauss III JF |
| Citation(s) |
17148555 |
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| Submission date |
Sep 15, 2006 |
| Last update date |
Mar 25, 2019 |
| Contact name |
Jennifer R Wood |
| E-mail(s) |
jwood5@unl.edu
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| Phone |
402-472-6437
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| Fax |
402-472-6362
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| Organization name |
University of Nebraska
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| Department |
Animal Science
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| Lab |
Wood
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| Street address |
3800 Fair St.
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| City |
Lincoln |
| State/province |
NE |
| ZIP/Postal code |
68583-0908 |
| Country |
USA |
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| Platforms (1) |
| GPL570 |
[HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array |
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| Samples (12)
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| GSM136511 |
Microarray analysis of NL and PCOS oocytes (NL1) |
| GSM136512 |
Microarray analysis of NL and PCOS oocytes (NL2) |
| GSM136513 |
Microarray analysis of NL and PCOS oocytes (NL3) |
| GSM136519 |
Microarray analysis of NL and PCOS oocytes (NL4) |
| GSM136525 |
Microarray analysis of NL and PCOS oocytes (NL5) |
| GSM136526 |
Microarray analysis of NL and PCOS oocytes (NL6) |
| GSM136527 |
Microarray analysis of NL and PCOS oocytes (P1) |
| GSM136533 |
Microarray analysis of NL and PCOS oocytes (P2) |
| GSM136554 |
Microarray analysis of NL and PCOS oocytes (P3) |
| GSM136560 |
Microarray analysis of NL and PCOS oocytes (P4) |
| GSM136561 |
Microarray analysis of NL and PCOS oocytes (P5) |
| GSM136562 |
Microarray analysis of NL and PCOS oocytes (P6) |
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| Relations |
| BioProject |
PRJNA97255 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE5850_RAW.tar |
88.6 Mb |
(http)(custom) |
TAR (of CEL, EXP) |
| Processed data included within Sample table |
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