title: Epithelial cell transforming protein 2 (ECT2) depletion blocks polar body extrusion and generates mouse oocytes containing two metaphase II spindles.
abstract: Completion of the first meiosis in oocytes is achieved by the extrusion of the first polar body (PBI), a particular example of cell division. In mitosis, the small GTPase RhoA, which is activated by epithelial cell transforming protein 2 (ECT2), orchestrates contractile ring constriction, thus enabling cytokinesis. However, the involvement of this pathway in mammalian oocytes has not been established. To characterize the role of ECT2 in PBI emission in mouse oocytes, the small interfering RNA approach was employed. We found that ECT2 depletion significantly reduces PBI emission, induces first metaphase arrest, and generates oocytes containing two properly formed spindles of the second metaphase. Moreover, we describe, for the first time, that before PBI emission, RhoA forms a ring that is preceded by a dome-like accumulation at the oocyte cortex, next to the spindle. This unique mode of RhoA translocation failed to occur in the absence of ECT2. We further found that the Rho-dependent kinase, a main RhoA effector, is essential for PBI emission. In addition, we demonstrate herein that ECT2 is subjected to phosphorylation/dephosphorylation throughout meiosis in oocytes and further reveal that PBI emission is temporally associated with ECT2 dephosphorylation. Our data provide the first demonstration that an active cyclin-dependent kinase 1, the catalytic subunit of the maturation-promoting factor, phosphorylates ECT2 during the first meiotic metaphase and that cyclin-dependent kinase 1 inactivation at anaphase allows ECT2 dephosphorylation. In conclusion, our study demonstrates the indispensable role of the maturation-promoting factor/ECT2/RhoA pathway in PBI extrusion in mouse oocytes.
authors.affiliation: Weizmann Institute of Science, Department of Biological Regulation, Herzel Street 1, Rehovot 76100, Israel.
authors.forename: Judith
authors.initials: J
authors.lastname: Elbaz
authors.forename: Yitzhak
authors.initials: Y
authors.lastname: Reizel
authors.forename: Nava
authors.initials: N
authors.lastname: Nevo
authors.forename: Dalia
authors.initials: D
authors.lastname: Galiani
authors.forename: Nava
authors.initials: N
authors.lastname: Dekel
mesh_terms: D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism; D018929:Cell Culture Techniques; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D006064:Gonadotropins, Equine / Q000494:pharmacology; D008677:Metaphase / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D006080:Ovarian Follicle / Q000187:drug effects / Q000502:physiology; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000172:deficiency* / Q000235:genetics / Q000502:physiology; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D034741:RNA, Small Interfering / Q000235:genetics; D008941:Spindle Apparatus / Q000502:physiology*; D020742:rhoA GTP-Binding Protein / Q000235:genetics
chemical_list: D018076:DNA, Complementary; C080509:Ect2 protein, mouse; D006064:Gonadotropins, Equine; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D012313:RNA; D016203:CDC2 Protein Kinase; D020742:rhoA GTP-Binding Protein