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ABSTRACT: Cumulus cells (CCs) are essential for oocytes to reach full development competency and become fertilized. Many major functional properties of CCs are triggered by gonadotropins and governed by the oocyte. Consequently, cumulus may reflect oocyte quality and is often used for oocyte selection. The most visible function of CCs is their ability for rapid extracellular matrix expansion after the LH surge. Although unexplained, LH induces the final maturation and improves oocyte quality. To study the LH signaling and gene expression cascade patterns close to the germinal vesicle breakdown, bovine CCs collected at 2 h before and 6 h after the LH surge were hybridized to a custom-made microarray to better understand the LH genomic action and find differentially expressed genes associated with the LH-induced oocyte final maturation. Functional genomic analysis of the 141 overexpressed and 161 underexpressed clones was performed according to their molecular functions, gene networks, and cell compartments. Following real-time PCR validation of our gene lists, some interesting pathways associated with the LH genomic action on CCs and their possible roles in oocyte final maturation, ovulation, and fertilization are discussed. A list of early potential markers of oocyte competency in vivo and in vitro is thereafter suggested. These early biomarkers are a preamble to understand the LH molecular pathways that trigger the final oocyte competence acquisition process in bovine.
Reproduction 12/2010; 140(6):835-52. · 2.58 Impact Factor
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ABSTRACT: To identify reliable genomic biomarkers expressed in cumulus cells that accurately and non-invasively predict the oocyte developmental competence and reinforce the already used morphological criteria.
Eight consenting patients were selected for ovarian stimulation and ICSI procedures. Cumulus-oocyte complexes were transvaginally punctured and individually selected based on both good morphological criteria and high zona pellucida birefringence. Following ICSI, two 3-day embryos per patient were transferred. Pregnancy outcome was recorded and proven implantation was thereafter confirmed. Differential gene expression was assessed using two microarray platforms. Further real-time PCR validation, Ingenuity pathways analysis and intra-patient analysis were performed on 17 selected candidates.
Seven genes were differentially (p ≤ 0.05) associated to successful pregnancy and implantation. These biomarkers could be used to predict the oocyte developmental competence.
These genomic markers are a powerful reinforcement of morphological approaches of oocyte selection. Their large-scale validation could increase pregnancy outcome and single embryo transfer efficiency.
Journal of Assisted Reproduction and Genetics 10/2010; 28(2):173-88. · 1.84 Impact Factor
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ABSTRACT: Oocyte competence is the ability of the oocyte to complete maturation, undergo successful fertilization, and reach the blastocyst stage. Cumulus cells are indispensable for this process. Their removal significantly affects the blastocyst rates. Moreover, the properties and functions of cumulus cells are regulated by the oocyte. They also reflect the oocyte's degree of maturation. Our study was aimed at identifying markers of oocyte competence that are expressed in bovine cumulus cells. In a previous study in our laboratory, the blastocyst yield following FSH or phorbol myristate acetate (PMA) treatment was 45%%. Therefore, we tested four sets of conditions during the first 6 h of in vitro maturation (IVM): FSH (0.1 microg/ml), PMA (0.1 microM), FSH ++ PMA, and negative control. Extracts from each IVM treatment were hybridized against the same negative control on a microarray containing a partial library of differentially expressed transcripts in the cumulus of competent oocytes collected at 6 h after LH in vivo. Common positive clones between diffrentially treated cells were selected, and 15 candidates were validated by real-time PCR. Based on this, the main candidates expressed in cumulus cells and that could be valuable and indirect markers of oocyte competence are hyaluronan synthase 2 (HAS2), inhibin betaA (INHBA), epidermal growth factor receptor (EGFR), gremlin 1 (GREM1), betacellulin (BTC), CD44, tumor necrosis factor-induced protein 6 (TNFAIP6), and prostaglandin-endoperoxide synthase 2 (PTGS2). These biomarkers could be potential candidates to predict oocyte competence and to select higher-quality embryos for transfer. Additionally, these indirect predictors of oocyte competence and follicular health could improve our knowledge of gene expression patterns in the cumulus and yield insights into the molecular pathways controlling oocyte competence.
Biology of Reproduction 05/2008; 79(2):209-22. · 4.01 Impact Factor
