Calcineurin (CN) is a serine/threonine phosphatase which plays important roles in meiosis maturation in invertebrate oocytes; however, the role of CN in mouse oocytes is relatively unexplored. In this study, we examined the expression, localization and functional roles of CN in mouse oocytes and granulosa cells. The RT-PCR results showed that the β isoform of calcineurin A subunit (Cn A) expres...

The requirement of mitogen-activated protein kinase (MAPK) for the initiation of meiotic maturation in porcine oocytes was examined using U0126, an inhibitor of MAPK activator. Immunoblot analysis and kinase assay showed that U0126 inhibited the phosphorylation and activation of MAPK incompletely in porcine oocytes. In this condition, maturation/M-phase promoting factor (MPF) activity was also ...

This study examines the timing of changes in chromosome configurations of pig oocytes derived from small antral follicles of follicular and/or inactive stage donors using modified 199 medium supplemented with gonadotropins (Follicle Stimulating Hormone (FSH), 10 IU/ml; Human Chorionic Gonadotropin (hCG), 10 IU/ml) and Glucosamine (0.539 mg/ml). Oocytes (n = 1,215) were fixed at the end of 3 hou...

As a tumor suppressor homologue during mitosis, Chk2 is involved in replication checkpoints, DNA repair, and cell cycle arrest, although its functions during mouse oocyte meiosis and early embryo development remain uncertain. We investigated the functions of Chk2 during mouse oocyte maturation and early embryo development. Chk2 exhibited a dynamic localization pattern; Chk2 expression was restr...

Developing a culture system for preantral follicles has important biotechnological implications due to the potential to produce a large number of oocytes for embryo production and transfer. To accomplish this goal, the present study was aimed to culture preantral follicles in the presence of different media, sera and FSH concentrations. Six-week-old preantral follicles (95 ± 5 µm) were cultured...

Total inhibition of germinal vesicle breakdown (GVBD) was observed in growing pig oocytes (internal diameter 80, 90 and 100 microns) when they were cultured in a medium conditioned by cumulus oocyte complexes (COCs) of fully grown oocytes. In denuded growing oocytes, only partial inhibition was observed. The inhibitory effect was fully reversible. The addition of heparin (300 IU/ml) could overc...

Tosti, Elisabetta, Raffaele Boni, and Annunziata Cuomo. Ca current activity decreases during meiotic progression in bovine oocytes. Am J Physiol Cell Physiol 279: C1795–C1800, 2000.—By using the whole cell voltage-clamp technique, we studied changes in plasma membrane permeability at different meiotic stages of bovine oocytes. Follicular oocytes were matured in vitro and activated by Ca ionopho...

The SUN (Sad-1/UNC-84) and KASH (Klarsicht/ANC-1/Syne/homology) proteins constitute the linker of nucleoskeleton and cytoskeleton (LINC) complex on the nuclear envelope. To date, the SUN1/KASH5 complex is known to function as meiotic-specific factors. In this study, gene-silencing methods were used to explore the roles of SUN1 and KASH5 in mouse oocytes after prophase. SUN1 was detected through...

Porcine cumulus oocyte complexes (COCs) were cultured together in 10-microliters droplets of culture medium. When 10 COCs were cultured for 24 h, germinal vesicle breakdown (GVBD) occurred in 81% of them. When more COCs (20 or 40) were put into the same volume of medium the frequency of GVBD gradually decreased. This inhibition was not observed in denuded oocytes. The process of GVBD was advers...

Efforts to improve the quality of in vitro matured oocytes by blocking germinal vesicle breakdown (GVBD) and allowing more time for ooplasmic maturation have achieved little due to a lack of knowledge on the molecular events during GVBD blocking. Such knowledge is also important for studies aimed at regulating gene expression in maturing oocytes prior to GVBD. We studied species difference and ...