Effects of RU486 and indomethacin on meiotic maturation, formation of extracellular matrix, and progesterone production by porcine oocyte-cumulus complexes

• Published in: Domestic animal endocrinology 2014-07, Vol.48, p.7-14
• Main Authors: Nagyova, E., Scsukova, S., Kalous, J., Mlynarcikova, A.
• Format: Article
• Language: English
• Subjects: Animals; C-Reactive Protein - genetics; C-Reactive Protein - metabolism; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - metabolism; Cumulus Cells - drug effects; Cumulus Cells - physiology; Cyclooxygenase Inhibitors - pharmacology; Extracellular Matrix - metabolism; Follicle Stimulating Hormone; Gene Expression Regulation - drug effects; Granulosa; Hormone Antagonists - pharmacology; Hyaluronic Acid; In Vitro Oocyte Maturation Techniques - veterinary; Indomethacin; Indomethacin - pharmacology; Luteinizing Hormone; Mifepristone - pharmacology; Oocyte; Oocytes - drug effects; Oocytes - physiology; Progesterone; Progesterone - metabolism; RU486; Serum Amyloid P-Component - genetics; Serum Amyloid P-Component - metabolism; Swine
• ISSN: 0739-7240; 1879-0054
• DOI: 10.1016/j.domaniend.2014.01.003

This study was designed to determine whether inhibition of either cyclooxygenase-2 (COX-2) by indomethacin or progesterone receptor (PR) by PR antagonist, RU486, affects oocyte maturation, progesterone production, and covalent binding between hyaluronan (HA) and heavy chains of inter-α trypsin inhibitor, as well as expression of cumulus expansion-associated proteins (HA-binding protein, tumor necrosis factor α-induced protein 6, pentraxin 3) in oocyte-cumulus complexes (OCCs). The experiments were based on freshly isolated porcine OCC cultures in which the consequences of PR and COX-2 inhibition on the final processes of oocyte maturation were determined. Granulosa cells (GCs) and OCCs were cultured in medium supplemented with FSH/LH (both 100 ng/mL) in the presence/absence of RU486 or indomethacin. Western blot analysis, 3H-glucosamine hydrochloride assay, immunofluorescence, and radioimmunoassay were performed. Only treatment with RU486 (25 μM) caused a decrease in the number of oocytes that reached germinal vesicle breakdown and metaphase II stage compared with indomethacin (100 μM) or FSH/LH treatment alone after 44 h. All treated OCCs synthesized an almost equal amount of HA. Heavy chains (of inter-α trypsin inhibitor)-HA covalent complexes were formed during in vitro FSH/LH-stimulated expansion in RU486- or indomethacin-treated OCCs. Follicle-stimulating hormone/LH-induced progesterone production by OCCs was increased in the presence of RU486 after 44 h. In contrast, a decrease of FSH/LH-stimulated progesterone production by GCs was detected in the presence of either RU486 or indomethacin after 72 h. We suggest that the PR-dependent pathway may be involved in the regulation of oocyte maturation. Both PR and COX-2 regulate FSH/LH-stimulated progesterone production by OCCs and GCs.