title: Meiosis-activating sterol-mediated resumption of meiosis in mouse oocytes in vitro is influenced by protein synthesis inhibition and cholera toxin.
abstract: To explore the possible signaling pathways of meiosis-activating sterol (MAS)-induced oocyte maturation and to elucidate whether the MAS pathway involves transcription or translation, arrested immature mouse oocytes were cultured with either the protein synthesis inhibitor cycloheximide or the heteronuclear RNA inhibitors alpha-amanitin or actinomycin D, respectively. Moreover, the possible involvement of a G protein-coupled receptor mechanism in MAS-mediated oocyte maturation was explored by influencing oocyte maturation with cholera toxin (CT). MAS-induced oocyte maturation was completely blocked by the addition of 50 microg/ml cycloheximide 4 h before the addition of MAS. Simultaneous addition of MAS and the protein synthesis inhibitor also significantly reduced the meiotic resumption compared to that in MAS-treated controls. In contrast, neither of the treatment regimens to inhibit transcription of DNA to RNA was observed to have any effect on the MAS-induced resumption of meiosis. CT was observed to inhibit MAS-induced, but not spontaneous, oocyte maturation in vitro, suggesting a putative involvement of G protein-coupled receptor mechanism in the MAS mode of action. In conclusion, protein synthesis was found to be an essential requirement for maintaining the oocytes' responsiveness to MAS-induced resumption of meiosis, in contrast to transcription.
authors.affiliation: Health Care Discovery, Pharmacology, Novo Nordisk A/S, Copenhagen, Denmark. Research Laboratories, Schering AG, Berlin, Germany. chgr@novo.dk
authors.forename: C
authors.initials: C
authors.lastname: Grøndahl
authors.forename: M
authors.initials: M
authors.lastname: Lessl
authors.forename: I
authors.initials: I
authors.lastname: Faerge
authors.forename: C
authors.initials: C
authors.lastname: Hegele-Hartung
authors.forename: K
authors.initials: K
authors.lastname: Wassermann
authors.forename: J L
authors.initials: JL
authors.lastname: Ottesen
mesh_terms: D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002772:Cholera Toxin / Q000494:pharmacology*; D002782:Cholestenes / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D005260:Female; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000378:metabolism; D008540:Meiosis; D051379:Mice; D008815:Mice, Inbred Strains; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D015398:Signal Transduction; D013261:Sterols / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D000546:Amanitins; D002782:Cholestenes; D019384:Nucleic Acid Synthesis Inhibitors; D011500:Protein Synthesis Inhibitors; D013261:Sterols; D003609:Dactinomycin; C100324:4,4-dimethylcholesta-8,14,24-trienol; D002772:Cholera Toxin; D003513:Cycloheximide; D019204:GTP-Binding Proteins