title: Effects of RU486 and indomethacin on meiotic maturation, formation of extracellular matrix, and progesterone production by porcine oocyte-cumulus complexes.
abstract: 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, (3)H-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.
authors.affiliation: Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, 27721 Libechov, Czech Republic. Electronic address: nagyova@iapg.cas.cz.
authors.forename: E
authors.initials: E
authors.lastname: Nagyova
authors.affiliation: Institute of Experimental Endocrinology, Slovak Academy of Sciences, 83301 Bratislava, Slovakia.
authors.forename: S
authors.initials: S
authors.lastname: Scsukova
authors.affiliation: Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, 27721 Libechov, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Kalous
authors.affiliation: Institute of Experimental Endocrinology, Slovak Academy of Sciences, 83301 Bratislava, Slovakia.
authors.forename: A
authors.initials: A
authors.lastname: Mlynarcikova
mesh_terms: D000818:Animals; D002097:C-Reactive Protein / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D054885:Cumulus Cells / Q000187:drug effects* / Q000502:physiology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D005640:Follicle Stimulating Hormone; D005786:Gene Expression Regulation / Q000187:drug effects; D006727:Hormone Antagonists / Q000494:pharmacology; D006820:Hyaluronic Acid; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary; D007213:Indomethacin / Q000494:pharmacology*; D007986:Luteinizing Hormone; D015735:Mifepristone / Q000494:pharmacology*; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D011374:Progesterone / Q000378:metabolism*; D000683:Serum Amyloid P-Component / Q000235:genetics / Q000378:metabolism; D013552:Swine; D000099153:Pentraxins
chemical_list: D002097:C-Reactive Protein; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D016861:Cyclooxygenase Inhibitors; D005640:Follicle Stimulating Hormone; D006727:Hormone Antagonists; D006820:Hyaluronic Acid; D007213:Indomethacin; D007986:Luteinizing Hormone; D015735:Mifepristone; D011374:Progesterone; D000683:Serum Amyloid P-Component; D000099153:Pentraxins
keywords: Granulosa; Indomethacin; Oocyte; Progesterone; RU486