Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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The transitional Gene Expression Profiling of Ovarian Follicle Treated with Indomethacin and RU486 in Rats

ABSTRACT: Indomethacin (an inhibitor of prostaglandin synthesis) and RU486 (mifepristone, progesterone receptor antagonist) are well known compounds which impair follicle rupture in rats, nevertheless the histopahotlogical figures of these follicle were different in detail due to the difference of the pathway which each compound inhibits. To characterize these unruptured follicles further and to investigate the genes which were altered in each rat treated with indomethacin or mifepristone, gene expression profile in ovarian follicle were analyzed in female sprague-Dawley rats. Peri- or post ovulatory follicles were collected by laser microdissection and extracted RNA was analyzed by genechip after single dosage of mifepristone or indomethacin. Animals were singly administered orally Indomethacin (IM) at 4mg/kg at 16:00 or RU486 (RU) at 100 mg/kg at 10:00 on the proestrus day, and sacrificed by exsanguination under anesthesia at 22:00 on the proestrus day (PeF: peri-ovulatory follicle), and 10:00 on the estrus day (PoF: post-ovulatory follicle). Ovaries were also collected from untreated rats at 22:00 on the proestrus day, and 10:00 on the estrus day as control groups. Three animals were contained per each treatment group at each timing. The ovaries were removed and embedded in OCT (Tissue-Tek, Sakura Finetek, Torrance, CA), frozen in dry-ice-cold isopentan, and stored at -70 C until used. Frozen sections (8 micro meter thick) were mounted on the membrane slides (MMI, Glatteburg, Zurich, Switzerland), and stained with HistogeneTM LCM frozen section staining kit (Arcturus Engineering, Mountain View, CA). Then, 15 follicles in each phase were retrieved by Laser Micro Dissection system (LMD, CellCut Plus, MMI). The collected follicle sections were lysed by the buffer RLT in the collection tube supplied in RNeasy Micro Kit (Qiagen, Hilden, Germany), and total RNA was extracted according to manufacturer's instructions.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Kenjiro Tsubota

PROVIDER: E-GEOD-54584 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Altered gene expression profile in ovarian follicle in rats treated with indomethacin and RU486.

Tsubota Kenjiro K Kanki Masayuki M Noto Takahisa T Nakatsuji Shunji S Oishi Yuji Y Matsumoto Masahiro M Nakayama Hiroyuki H

The Journal of toxicological sciences 20150601 3

It is well-known that indomethacin (the cyclooxygenase 1 & 2 inhibitor) and RU486 (or mifepristone, the progesterone receptor antagonist) block follicular rupture in rats. To characterize genetic alterations in unruptured follicles, gene expression profiles in ovarian follicle were analyzed in indomethacin- and RU486-treated female Sprague-Dawley rats. Ovaries are collected at 22:00 on the proestrus day and 10:00 on the following estrus day after a single dose of indomethacin and RU486. Histopat ...[more]

PMID: 25972201

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Project description:Experiments were designed to evaluate changes in the transcriptome (mRNA levels) in the ovulatory, luteinizing follicle of rhesus monkeys, using a controlled ovulation (COv) model that permits analysis of the naturally selected, dominant follicle at specific intervals (0, 12, 24, 36 hours) after exposure to an ovulatory (exogenous hCG) stimulus during the menstrual cycle. Total RNA was prepared from individual follicles (n=4-8/timepoint), with an aliquot used for microarray analysis (AffymetrixTM Rhesus Macaque Genome Array) and the remainder applied to quantitative real-time PCR (q-PCR) assays. The microarray data from individual samples distinctly clustered according to timepoints, and ovulated follicles displayed markedly different expression patterns from unruptured follicles at 36 h. Between timepoint comparisons revealed profound changes in mRNA expression profiles. The dynamic pattern of mRNA expression for steroidogenic enzymes (CYP17A, CYP19A, HSD3B2, HSD11B1, HSD11B2), StAR, and gonadotropin receptors (LHCGR, FSHR) as determined by microarray analysis correlated precisely with those from blinded q-PCR assays. Patterns of mRNA expression for EGF-like factors (AREG, EREG) and processes (HAS2, TNFAIP6) implicated in cumulus-oocyte maturation/expansion were also comparable between assays. Thus, several mRNAs displayed the expected expression pattern for purported theca (e.g., CYP17A, AREG), granulosa (CYP19A, FSHR), cumulus (HAS2, TNFAIP6) cell, and surface epithelium (HSD11B) related genes in the rodent/primate preovulatory follicle. This database will be of great value in analyzing molecular and cellular pathways associated with periovulatory events in the primate follicle (e.g. follicle rupture, luteinization, inflammatory response, and angiogenesis), and for identifying novel gene products controlling mammalian fertility. Keywords: time course Total RNA was prepared from individual follicles (n=4-8/timepoint). Dominant follicles were selected specific intervals (0, 12, 24, 36 hours) after exposure to an ovulatory (exogenous hCG) stimulus during the menstrual cycle.

Dataset Information

The transitional Gene Expression Profiling of Ovarian Follicle Treated with Indomethacin and RU486 in Rats

Publications

Altered gene expression profile in ovarian follicle in rats treated with indomethacin and RU486.

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