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钙刺激后牛卵母细胞中的组蛋白H1激酶活性。

Histone H1 kinase activity in bovine oocytes following calcium stimulation.

作者信息

Collas P, Sullivan E J, Barnes F L

机构信息

GenMark, Inc., Salt Lake City, Utah 84108.

出版信息

Mol Reprod Dev. 1993 Feb;34(2):224-31. doi: 10.1002/mrd.1080340215.

Abstract

The influence of number of Ca2+ stimulations on the profile of histone H1 kinase activity in bovine oocytes was investigated. A CA2+ stimulation consisted of transferring oocytes directly from culture medium to mannitol containing 100 microM Ca2+ and pulsing oocytes with a 0.2 kVcm-1, 20 microseconds discharge. One, three, or six Ca2+ stimulations were given, each 22 min apart. Oocytes were frozen from 0 to 8 hr after the first stimulation at indicated time points and assayed for histone H1 kinase activity. H1 kinase activity was quantified using a densitometer and expressed as a percent of activity in nonpulsed metaphase II oocytes. Stimulating oocytes in the absence of Ca2+ in the pulsing medium did not inactivate H1 kinase. In the presence of Ca2+, however, H1 kinase was rapidly inactivated after stimulation. A single stimulation decreased H1 kinase activity to 44% +/- 11% of its initial level in 1 hr. However, H1 kinase was dramatically reactivated at 2 hr after the stimulation and reached 122% +/- 22% of the initial activity at 6 hr. With three stimulations, basal H1 kinase activity was 21% +/- 3% and was obtained in 30 min. H1 kinase reactivation started at 4 hr after the first stimulation and level of activity reached 38% +/- 15% at 8 hr. Six stimulations also led to rapid H1 kinase inactivation and to a basal activity of 14% +/- 0.4%. With six stimulations, however, basal H1 kinase activity was maintained over at least 8 hr, and no reactivation occurred during this period. Basal H1 kinase activity obtained after six stimulations was similar to that of fertilized oocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

研究了Ca2+刺激次数对牛卵母细胞组蛋白H1激酶活性曲线的影响。一次Ca2+刺激包括将卵母细胞直接从培养基转移至含100微摩尔Ca2+的甘露醇中,并用0.2千伏/厘米、20微秒的放电脉冲处理卵母细胞。分别给予1次、3次或6次Ca2+刺激,每次间隔22分钟。在首次刺激后的0至8小时内,于指定时间点将卵母细胞冻存,并检测组蛋白H1激酶活性。使用密度计对H1激酶活性进行定量,并表示为未脉冲处理的中期II期卵母细胞活性的百分比。在脉冲培养基中无Ca2+时刺激卵母细胞不会使H1激酶失活。然而,在有Ca2+存在的情况下,刺激后H1激酶会迅速失活。单次刺激在1小时内使H1激酶活性降至其初始水平的44%±11%。然而,H1激酶在刺激后2小时显著重新激活,并在6小时达到初始活性的122%±22%。给予3次刺激时,基础H1激酶活性为21%±3%,在30分钟时获得。H1激酶重新激活在首次刺激后4小时开始,8小时时活性水平达到38%±15%。6次刺激也导致H1激酶迅速失活,基础活性为14%±0.4%。然而,给予6次刺激后,基础H1激酶活性至少维持8小时,在此期间未发生重新激活。6次刺激后获得的基础H1激酶活性与受精卵母细胞的相似。(摘要截短至250字)

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