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Chromatin Spread Preparations for the Analysis of Mouse Oocyte Progression from Prophase to Metaphase II
Published on: February 26, 2018
Meiosis resumption, calcium-sensitive period, and PLC-beta1 relocation into the nucleus in the mouse oocyte
Nathalie Avazeri1, Anne-Marie Courtot, Arlette Pesty
1Institut National de la Santé et de la Recherche Médicale Unité 566, CEA/DSV/LMMG Bâtiment 05 BP6, 92265 Fontenay-aux-Roses, France.
Abstract:
We aimed to determine whether the breakdown of the germinal vesicle of the mouse oocyte and the nuclear import of phospholipase C-beta1 were calcium-dependent. We chelated Ca2+ ions with BAPTA-dextran at different times after the release of the oocyte from the ovarian follicle, i.e. after meiosis resumption has started, and we studied the effects on the kinetics of germinal vesicle breakdown, and on the migration of phospholipase C-beta1. We discriminate between two key-periods of calcium-sensitivity during the process of meiosis resumption. During the first hour, changes in the cytosolic Ca2+ especially promoted the migration of phospholipase C-beta1 into the nucleus, whereas changes in the nuclear concentration of Ca2+ were not implicated. Moreover, at this time, the cytosolic calcium pathway is PLC-beta1-dependent. By contrast, during the second hour following the onset of meiosis resumption, and thus just previous GVBD, the PLC-beta1-dependent Ca2+ signals in both cellular compartments were equally necessary for the resumption of meiosis. This particular period of the meiotic process corresponds to the moment when the phospholipase C-beta1 has strongly migrated into the nucleus. Our results highlight also the role played by the nucleus during the second key-period in the control of the GVBD via a Ca2+-dependent pathway.
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