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Increased parthenogenetic development of bovine and porcine oocytes activated with a combination of a protein synthesis and phosphorylation inhibitor.

机译:通过蛋白质合成和磷酸化抑制剂的组合激活的牛和猪卵母细胞的孤雌生殖发育增加。

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摘要

Calcium ionophores such as A23187 (A) are used to artificially trigger the release of free intracellular Ca2+ within oocytes. Calcium ionophores cannot effectively induce parthenogenetic development in bovine and porcine oocytes alone. Therefore, it is necessary to combine the ionophore with either a protein synthesis inhibitor or a phosphorylation inhibitor to initiate development. Cycloheximide (CHX), maintains low MPF activity in the oocyte by inhibiting cyclin B synthesis. Exposing oocytes to 6-dimethylaminopurine (6-DMAP) induces early inactivation of MAPK through the dephosphorylation of ERK2. Regulating these proteins is critical as they are responsible for the resumption of meiosis and microtubule/chromatin organization. These studies were designed to compare the cell cycle progression and development of bovine and porcine oocytes activated with CHX and 6-DMAP alone or in combination with each other. Oocytes treated with the triple chemical combination of A23187, CHX and 6-DMAP had significantly higher rates of blastocyst development compared to conventional activation methods. It is possible that the combination of a protein synthesis inhibitor and a phosphorylation inhibitor better mimics natural fertilization by inactivating both MPF and MAPK. Results from this study may be beneficial to other areas of biotechnology such as cloning in which chemical activation is required for development.
机译:钙离子载体(例如A23187(A))用于人工触发卵母细胞中游离细胞内Ca2 +的释放。钙离子载体不能仅在牛和猪卵母细胞中有效诱导孤雌生殖发育。因此,有必要将离子载体与蛋白质合成抑制剂或磷酸化抑制剂结合以启动显影。环己酰亚胺(CHX)通过抑制细胞周期蛋白B的合成,在卵母细胞中维持低MPF活性。将卵母细胞暴露于6-二甲基氨基嘌呤(6-DMAP)通过ERK2的去磷酸化诱导MAPK的早期失活。调节这些蛋白质至关重要,因为它们负责恢复减数分裂和微管/染色质的组织。设计这些研究的目的是比较单独或联合使用CHX和6-DMAP激活的牛和猪卵母细胞的细胞周期进程和发育。与传统的激活方法相比,用A23187,CHX和6-DMAP的三重化学组合处理的卵母细胞的胚泡发育速率明显更高。蛋白质合成抑制剂和磷酸化抑制剂的组合可能通过使MPF和MAPK失活而更好地模拟自然受精。这项研究的结果可能有益于生物技术的其他领域,例如需要化学激活才能进行开发的克隆。

著录项

  • 作者

    Bormann, Charles Louis.;

  • 作者单位

    University of Connecticut.;

  • 授予单位 University of Connecticut.;
  • 学科 Biology Physiology.
  • 学位 Ph.D.
  • 年度 2006
  • 页码 131 p.
  • 总页数 131
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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