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In Vitro Maturation and Parthenogenetic Activation of Bovine and Porcine Oocytes

牛和猪卵母细胞的体外成熟和孤雌激活

基本信息

批准号：
10660263
负责人：
SUZUKI HIROYUKI
金额：
$ 2.18万
依托单位：
HIROSAKI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660263/
关键词：
Cattle Pi Oocytes Activation SEM Zona pellucida Microtubules Microfilaments ウシ卵母細胞表面微細構造

项目摘要

Parthenogenetic activation of oocytes is a key component of the current mammalian cloning regimen. This study was designed to examine the cellular alterations of the bovine and porcine oocytes at different ages following different activation stimulations. In Experiments 1 and 2, bovine oocytes of 22,30 and 40 h after in vitro maturation was activated by an electrical pulse (100Vmm, 50μsec), Ca ionophore, and combined with cycloheximide or 6-dimethylaminopurine. It is suggested that older bovine oocytes are more readily activated and that the age-dependent surface ultrastructural alterations correlate very well with the age dependent activation of oocytes. It was also shown that surface characteristics of the bovine oocyte were more profoundly changed by a combination of parthenogenetic stimulation, and that the ultrastructural effects were reversible. In Experiments 3 and 4, porcine oocytes were matured in vitro to examine morphodynamics of the cumulus-oocyte association during oocyte … More maturation. It was concluded that the cumulus-oocyte transzonal projections became disconnected between MI and MII stage as a result of cumulus expansion. However, the cumulus-cumulus communications remained intact at these stages. In Experiments 5 and 6, the activation rate, developmental pattern and changes in microtubule and microfilament assembly of the porcine oocytes activated electrically or chemically were evaluated in relation to the oocyte aging. The present results suggest that a higher activation rate was obtained in the porcine oocytes after two electrical pulses that in those after Ca ionophore or ethanol treatments and that the disorganization of microtubule and the alteration of microfilament distribution observed in the aged oocyte may result in higher incidences of diploid parthenotes and fragmentation, respectively. It is also suggested that increased development to blastocysts after the combined treatment of electrical pulses and CD may due to higher incidence of diploid parthenotes derived from CD treatment. Less

卵母细胞的孤雌激活是当前哺乳动物克隆方案的关键组成部分。本研究旨在检查不同年龄的牛和猪卵母细胞在不同激活刺激后的细胞变化。在实验1和2中，体外成熟22，30和40小时的牛卵母细胞通过电脉冲（100Vmm，50μsec）、Ca离子载体并与放线菌酮或6-二甲氨基嘌呤组合激活。这表明，年龄较大的牛卵母细胞更容易被激活，并且年龄依赖性表面超微结构的改变与卵母细胞年龄依赖性激活密切相关。研究还表明，孤雌生殖刺激的组合可以更深刻地改变牛卵母细胞的表面特征，并且超微结构效应是可逆的。在实验 3 和 4 中，猪卵母细胞在体外成熟，以检查卵母细胞成熟过程中卵丘-卵母细胞关联的形态动力学。结论是，由于卵丘扩张，卵丘-卵母细胞跨区投射在 MI 和 MII 阶段之间变得不连贯。然而，积云与积云之间的通讯在这些阶段保持完整。在实验5和6中，评估了电或化学激活的猪卵母细胞的激活率、发育模式以及微管和微丝组装的变化与卵母细胞老化的关系。目前的结果表明，两次电脉冲后猪卵母细胞比钙离子载体或乙醇处理后的卵母细胞获得了更高的活化率，并且在老化卵母细胞中观察到的微管解体和微丝分布的改变可能分别导致二倍体单性体和碎片的发生率更高。还表明，电脉冲和 CD 联合处理后囊胚发育增加可能是由于 CD 处理产生的二倍体单性动物的发生率较高。较少的