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

卵母细胞的孤态激活是当前哺乳动物克隆方案的关键组成部分。这项研究旨在检查不同活化刺激后不同年龄的牛和猪卵母细胞的细胞改变。在实验1和2中，体外成熟后22,30和40小时的牛卵母细胞通过电脉冲（100Vmm，50μsec），离子载体激活，并与环己酰胺或6-二甲基氨基肽结合使用。据建议，较老的牛卵母细胞更容易激活，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非常相关。还表明，牛卵母细胞的表面特征通过孤态发生刺激的结合而更深刻地改变，并且超微结构效应是可逆的。在实验3和4中，将猪卵母细胞在体外成熟，以检查卵母细胞期间卵母细胞缔合的形态动力学……更成熟。得出的结论是，由于积云的扩张，育素素细胞的跨分子投影在MI和MII阶段之间断开了连接。但是，在这些阶段，库卢斯库鲁斯通信仍然完好无损。在实验5和6中，与卵母细胞衰老有关，对猪卵母细胞的激活率，发育模式以及微丝和微丝组装的变化进行了评估。目前的结果表明，在两种电脉冲之后，在猪卵母细胞中获得了更高的活化率，即在CA离子载体或乙醇处理后的脉冲中，微管的混乱和在老年卵母细胞中观察到的微丝分布的变化可能会导致较高的型型卵巢卵母细胞和较高的卵黄细胞的发生率。还提出，在电脉冲联合处理和CD之后，增加对胚泡的发育可能是由于CD处理中二倍体parthenotes的较高入射而引起的。较少的