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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。本研究旨在研究不同年龄牛和猪卵母细胞在不同激活刺激下的细胞变化。实验1和实验2分别用电脉冲（100Vmm, 50μsec）、Ca离子离子激活体外成熟22,30和40h的牛卵母细胞，并与环己亚胺或6-二甲氨基嘌呤结合。这表明，年龄较大的牛卵母细胞更容易被激活，并且年龄依赖性的表面超微结构改变与卵母细胞的年龄依赖性激活密切相关。单性生殖联合刺激对牛卵母细胞表面特征的影响更为深刻，超微结构的影响是可逆的。在实验3和4中，猪卵母细胞在体外成熟，以观察卵母细胞成熟过程中卵丘-卵母细胞结合的形态动力学。结果表明，在MI和MII阶段，由于积云的扩张，积云-卵母细胞的跨区投射断开。然而，在这些阶段，积云-积云之间的通信保持完整。实验5和实验6分别对电激活和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进行了研究。实验结果表明，2次电脉冲处理对猪卵母细胞的激活率高于钙离子载体和乙醇处理，衰老卵母细胞微管的紊乱和微丝分布的改变可能分别导致二倍体孤雌体和断裂的发生率更高。电脉冲和CD联合处理后囊胚发育加快可能是由于CD处理产生的二倍体孤雌体发生率较高。少