Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mouse ovaries from FORKO mutants animals

ABSTRACT: To understand consequences of loss of FSH receptor, we used genetically altered mouse mutants with deletion of this receptor. Follitropin Receptor Knockout (FORKO) mice present various developmental and age-dependent abnormalities such as infertility, senescence and increased tumor incidences in addition to other phenotypes. To explore why the atrophic ovaries of FORKO mice develop ovarian anomalies later in life, we used expression profiling studies to gain a more comprehensive view of genes that are misregulated. Experiment Overall Design: Mice were housed under controlled temperature and constant light (12 h of light, 12 h of darkness), with food and water provided ad libitum. The female mice used in this experiment were derived by breeding heterozygotes of the 129T2 svEmsJ background. Experiment Overall Design: For RNA extraction, both the ovaries from 8 month old mice were carefully dissected and pooled. Tissues were frozen in liquid nitrogen and homogenized in TRIZOL (Invitrogen, Burlington, Ontario) and processed in accordance with the manufacturerâs protocol. Experiment Overall Design: Affymetrix 39 K GeneChipÂ® Mouse Genome 430 2.0 Array has over 45000 probe sets which analyses 39000+ transcripts and variants from over 34000 well characterized mouse genes. A total of 1 Âµg of double-stranded cDNA was transcribed in vitro using the Bioarray High-Yield RNA Transcript Labeling Kit (Enzo, Farmingdale, NY) according to the manufacturer's instructions using biotinylated CTP and UTP (Enzo, Farmingdale, NY). Following 5-hr incubation at 37Â°C, the resultant biotin-labeled cRNA was purified with RNAeasy columns (Qiagen) and eluted in 40 Âµl of RNAse-free water. The concentration of biotin-labeled cRNA was determined by RNA bio-analyzer. Target cRNAs corresponding to either wild type ovary or FORKO ovarian tissue were hybridized to an individual GeneChip from an identical lot of Affymetrix 39 K GeneChipÂ® Mouse Genome 430 2.0 array for 16 hr. GeneChip arrays were washed and stained using antibody-mediated signal amplification and the Affymetrix Fluidics Station's standard Eukaryotic GE Wash 2 protocol, using Affymetrix equipment and protocols (Affymetrix, Santa Clara, CA).

ORGANISM(S): Mus musculus

SUBMITTER: JAYAPRAKASH ARAVINDAKSHAN

PROVIDER: E-GEOD-4323 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Differential expression of claudin family proteins in mouse ovarian serous papillary epithelial adenoma in aging FSH receptor-deficient mutants.

Aravindakshan Jayaprakash J Chen Xinlei X Sairam M Ram MR

Neoplasia (New York, N.Y.) 20061201 12

Ovarian cancer is a deadly disease with long latency. To understand the consequences of loss of follicle-stimulating hormone receptor (FSH-R) signaling and to explore why the atrophic and anovulatory ovaries of follitropin receptor knockout (FORKO) mice develop different types of ovarian tumors, including serous papillary epithelial adenoma later in life, we used mRNA expression profiling to gain a comprehensive view of misregulated genes. Using real-time quantitative reverse transcription-polym ...[more]

PMID: 17217615

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Project description:Mice that develop benign cartilage lesions due to overexpression of Gli2 in chondrocytes developed lesions similar to chondrosarcomas when also deficient in p53. Gli2 overexpression and p53 deficiency had opposing effects on chondrocyte differentiation, but had additive effects negatively regulating apoptosis. Regulation of Igfbp3 expression and IGF signaling by Gli and p53 integrated their effect on apoptosis. Treatment of human chondrosarcomas or fetal mouse limbs explants with IGFBP3 or by blocking IGF increased the apoptosis rate, and mice expressing Gli2 developed substantially fewer tumors when also deficient for Igf2. IGF signaling meditated apoptosis regulates the progression to malignant chondrosarcoma. Experiment Overall Design: The CH10T1/2 cell line was stably transfected with either the R150C variant PTHR1 present in enchondromas (Hopyan, S., Gokgoz, N., Poon, R., Gensure, R. C., Yu, C., Cole, W. G., Bell, R. S., Juppner, H., Andrulis, I. L., Wunder, J. S., and Alman, B. A. (2002). A mutant PTH/PTHrP type I receptor in enchondromatosis. Nat Genet 30, 306-310) or a wild type PTHR1, as previously reported (Mau, E., Whetstone, H., Yu, C., Hopyan, S., Wunder, J. S., and Alman, B. A. (2007). PTHrP regulates growth plate chondrocyte differentiation and proliferation in a Gli3 dependent manner utilizing hedgehog ligand dependent and independent mechanisms. Dev Biol 305, 28-39). Cells were grown overnight in serum free media and treated with either 10â?? 7 M PTHrP (Bachem, King of Prussia, Pennsylvania, USA), or carrier alone for one hour. RNA was isolated from the cells CH10T1/2 cells expressing either the R150C variant PTHR1 or wild type PTHR1. RNA was converted to double-stranded cDNA using Superscript (Gibco-Invitrogen) with a T7-(dT)24 primer, which was then transcribed to biotinylated complementary RNA (cRNA) by incorporating biotin-CTP and biotin-UTP using Enzo BioArray High Yield RNA labeling kit (Enzo Diagnostics, New York, NY). The cRNA labeling and hybridizations were then performed according to Affymetrix GeneChip Protocol (Affymetrix Inc., Santa Clara, CA). The chips were scanned for fluorescence signal detection. Affymetrix - GeneChipÂ® Mouse Genome 430A 2.0 Array was utilized.

Dataset Information

Transcription profiling of mouse ovaries from FORKO mutants animals

Publications

Differential expression of claudin family proteins in mouse ovarian serous papillary epithelial adenoma in aging FSH receptor-deficient mutants.

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