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类固醇对猪卵母细胞体外成熟抑制作用的特异性

Specificity of inhibition by steroids of porcine oocyte maturation in vitro.

作者信息

Richter J D, McGaughey R W

出版信息

J Exp Zool. 1979 Jul;209(1):81-90. doi: 10.1002/jez.1402090110.

Abstract

In order to examine the influence of several steroids on the process of oocyte maturation, denuded (adherent cumulus granulosa cells mechanically removed) and intact (cumulus granulosa cells left attached) porcine oocytes were cultured in the presence or absence of estradiol-17 beta, estradiol-17 alpha, testosterone, cortisol, progesterone, or the nonsteroidal estrogen diethyl stilbestrol (all at 10 microgram/ml) in defined medium that contained either BSA or dextran. Estradiol-17 beta was the only steroid to exert a significant inhibitory effect on the maturation of denuded oocytes, and did so only in BSA supplemented medium. The inhibition was reversible in that oocytes, cultured in steroid-free medium after initial culture in estradiol-17 beta medium, resumed meiotic maturation. Oocytes took up 3H-estradiol-17 beta in both media, although less radiolabel entered oocytes in BSA supplemented medium. The majority of label in the oocytes, when cultured with either medium, was not displaced by excess radioinert estradiol-17 beta or progesterone, nor were the oocytes saturated even when cultured in 10(-6) M estradiol-17 beta. Autoradiography of sectioned oocytes after culture in 3H-estradiol-17 beta has shown that there was no selective accumulation of silver grains over the germinal vesicle as was the case with granulosa cell nuclei. This observation suggests that estradiol-17 beta may not act at the level of the oocyte nucleus.

摘要

为了研究几种甾体激素对卵母细胞成熟过程的影响,将去除颗粒细胞(机械去除附着的卵丘颗粒细胞)的猪卵母细胞和完整的(保留附着的卵丘颗粒细胞)猪卵母细胞,在含有牛血清白蛋白(BSA)或葡聚糖的限定培养基中,于存在或不存在17β-雌二醇、17α-雌二醇、睾酮、皮质醇、孕酮或非甾体雌激素己烯雌酚(均为10微克/毫升)的条件下进行培养。17β-雌二醇是唯一对去除颗粒细胞的卵母细胞成熟有显著抑制作用的甾体激素,且仅在添加BSA的培养基中如此。这种抑制是可逆的,因为在17β-雌二醇培养基中初步培养后,在无甾体激素的培养基中培养的卵母细胞恢复了减数分裂成熟。两种培养基中的卵母细胞均摄取了3H-17β-雌二醇,尽管在添加BSA的培养基中进入卵母细胞的放射性标记较少。当用任一种培养基培养时,卵母细胞中的大部分标记物不会被过量的放射性惰性17β-雌二醇或孕酮取代,即使在10⁻⁶ M 17β-雌二醇中培养,卵母细胞也未饱和。在3H-17β-雌二醇中培养后的卵母细胞切片放射自显影显示,与颗粒细胞核的情况不同,生发泡上没有银粒的选择性积累。这一观察结果表明,17β-雌二醇可能不在卵母细胞核水平发挥作用。

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