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Effects of butyrolactone-I on GVBD in bovine oocytes and subsequent maturation, fertilization and development in vitro

机译:丁内酯-I对牛卵母细胞GVBD的影响以及体外成熟,受精和发育

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摘要

In Experiment 1, bovine cumulus oocyte complexes (COCs) were preincubated for 24 h in M199 supplemented with 25-100 muM BL-I, an inhibitor of cdc2 kinase activity. Half of the COCs were fixed to assess rates of germinal vesicle breakdown (GVBD), and the rest were cultured for a further 20 h without BL-I to investigate whether BL-1 affects their meiotic competence. In Experiment 2, COCs treated with BL-I for 24 h or 48 h were matured (IVM), fertilized (IVF) and then cultured in vitro (IVC) for 8 days. Oocytes exposed to 100 muM BL-I showed a significantly higher rate of germinal vesicle (GV) (86%) stage than the others (4-72%). Oocytes exposed to 100 muM BL-I for 48 h showed equally high GV (95%) stage and maturation (84%) rates compared to oocytes treated for 24 h and control oocytes with no treatment. However, they showed significantly lower normal fertilization (27%), cleavage (27%) and blastocyst formation (6%) rates than oocytes treated for 24 h (66, 69 and 42%, respectively) and control oocytes (65, 75 and 42%, respectively). The results indicate that BL-I can reversibly inhibit GVBD of bovine oocytes for at least 24 h but not for 48 h without compromising subsequent developmental competence after IVM-IVF-IVC.
机译:在实验1中,将牛卵丘卵母细胞复合物(COC)在补充了25-100μMBL-1的M199中预孵育24小时,该BL-1是cdc2激酶活性的抑制剂。固定了一半的COC以评估发芽囊泡分解(GVBD)的速率,其余的COC在没有BL-1的情况下再培养20小时,以研究BL-1是否影响它们的减数分裂能力。在实验2中,用BL-1处理24小时或48小时的COC成熟(IVM),受精(IVF),然后体外培养(IVC)8天。暴露于100μMBL-1的卵母细胞的生小泡(GV)分期(86%)的比率显着高于其他卵母细胞(4-72%)。与经过24小时处理的卵母细胞和未经处理的对照卵母细胞相比,暴露于100μMBL-1持续48 h的卵母细胞显示出同样高的GV(95%)分期和成熟率(84%)。然而,它们显示正常受精(27%),卵裂(27%)和胚泡形成(6%)率显着低于处理24小时的卵母细胞(分别为66%,69%和42%)和对照卵母细胞(65、75和70%)。分别为42%)。结果表明,BL-1可以可逆地抑制牛卵母细胞的GVBD至少24 h,但不能持续48 h,而不会损害IVM-IVF-IVC后的后续发育能力。

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