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牛蛙卵提取物中成熟促进因子(MPF)的振荡活性。

Oscillatory activity of maturation promoting factor (MPF) in extracts of Rana pipiens eggs.

作者信息

Masui Y

出版信息

J Exp Zool. 1982 Dec 30;224(3):389-99. doi: 10.1002/jez.1402240312.

Abstract

Dejellied Rana pipiens eggs were crushed by centrifugation and clear extracts were prepared by further centrifugation at 150,000 X G for 2 h. The extracts were kept at 0 degrees C for 2 to 3 weeks. At intervals during the storage period, 80-nl aliquots were injected into ovarian oocytes to assay maturation promoting factor (MPF). In most cases, MPF activity disappeared from the extracts on day 3 or 4, but reappeared on the following days and persisted 2 or 3 days at high levels before disappearing again. The cycle of MPF activity in extracts was repeated fairly regularly a few times during storage of the extracts. The average period of the oscillation was 5.05 + 1.25 days. If a single extract was assayed by oocytes from different frogs, all the assays exhibited exactly the same patterns of changes in MPF activity during storage of the extract. This indicates that the oscillations in MPF activity observed during storage of the extracts reflect changes in MPF activity intrinsic to the extracts, rather than fluctuations in the sensitivity of recipient oocytes to MPF. The reappearance of MPF activity in stored extracts was sensitive to Ca and dependent on Mg ions. The MPF activity that recurred periodically in cold-stored extracts was indistinguishable from that contained in fresh extracts, since oocytes induced to mature by the recurring MPF activity had chromosomes condensed to meiotic metaphase and were capable of cleavage when injected with a sperm suspension. The oscillatory behavior of MPF activity observed in this study may be comprehended on a conceptual basis by aid of a kinetic model.

摘要

将去胶的豹蛙卵进行离心破碎,然后通过在150,000×g下进一步离心2小时制备清亮提取物。提取物保存在0℃下2至3周。在储存期间每隔一段时间,将80 nl的等分试样注射到卵巢卵母细胞中以测定成熟促进因子(MPF)。在大多数情况下,MPF活性在第3天或第4天从提取物中消失,但在接下来的几天又重新出现,并在再次消失之前高水平持续2或3天。在提取物储存期间,提取物中MPF活性的循环相当规律地重复了几次。振荡的平均周期为5.05±1.25天。如果用来自不同青蛙的卵母细胞对单一提取物进行测定,所有测定在提取物储存期间均表现出完全相同的MPF活性变化模式。这表明在提取物储存期间观察到的MPF活性振荡反映了提取物内在的MPF活性变化,而不是受体卵母细胞对MPF敏感性的波动。储存提取物中MPF活性的重新出现对Ca敏感且依赖于Mg离子。在冷藏提取物中周期性重现的MPF活性与新鲜提取物中所含的MPF活性没有区别,因为由重现的MPF活性诱导成熟的卵母细胞具有凝聚至减数分裂中期的染色体,并且在注射精子悬浮液时能够进行卵裂。本研究中观察到的MPF活性的振荡行为可以借助动力学模型在概念基础上进行理解。

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