Park Sol Ji, Koo Ok Jae, Kwon Dae Kee, Gomez Ma Ninia Limas, Kang Jung Taek, Atikuzzaman Mohammad, Kim Su Jin, Jang Goo, Lee Byeong Chun
Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, 151-742, Korea.
Zygote. 2011 Feb;19(1):1-8. doi: 10.1017/S0967199410000134. Epub 2010 Jun 23.
Treatment with 6-dimethylaminopurine (6-DMAP) or demecolcine (DE) for several (at least 2) hours after artificial activation is known to improve in vitro development of porcine embryos. However, several reports have also shown that treatments with these chemicals induce apoptosis. The aim of this study was to find out whether short-term treatment with 6-DMAP and DE combined with electrical or thimerosal/dithiothreitol (Thi/DTT) activation had a beneficial effect on development of parthenogenetically activated porcine oocytes. We additionally treated embryos with 6-DMAP (2 mM) and/or DE (0.4 microg/ml) for a short time (40 min) after an electrical pulse (EP) or Thi/DTT. As a result, short-term treatment with 6-DMAP and DE successfully induced development of electrically or Thi/DTT-activated porcine parthenogenetic embryos with no significant difference in cleavage rate, blastocyst formation rate and total cell number compared with long-term treatment. To find optimal activation protocol, cleavage rate, blastocyst formation rate and total cell number were compared between EP and Thi/DTT treatments. Thi/DTT + 6-DMAP + DE showed significantly higher blastocyst formation rate (36.1 ± 3.5%) and total cell number (46.9 ± 1.0) than other groups (EP + 6-DMAP + DE, EP + Thi/DTT + 6-DMAP + DE: 23.3 ± 3.0%, 42.2 ± 1.1 and 17.2 ± 2.7%, 36.7 ± 1.5, respectively). In conclusion, this study demonstrates that short-term treatment with 6-DMAP and DE is as effective as the standard long-term treatment and Thi/DTT + 6-DMAP + DE exerts a synergistic effect.
已知在人工激活后用6-二甲基氨基嘌呤（6-DMAP）或秋水仙酰胺（DE）处理数小时（至少2小时）可改善猪胚胎的体外发育。然而，也有几份报告表明，用这些化学物质处理会诱导细胞凋亡。本研究的目的是确定6-DMAP和DE短期处理与电激活或硫柳汞/二硫苏糖醇（Thi/DTT）激活相结合是否对孤雌激活的猪卵母细胞发育有有益影响。我们在电脉冲（EP）或Thi/DTT处理后，再用6-DMAP（2 mM）和/或DE（0.4 μg/ml）对胚胎进行短时间（40分钟）处理。结果，与长期处理相比，6-DMAP和DE短期处理成功诱导了电激活或Thi/DTT激活的猪孤雌胚胎的发育，在卵裂率、囊胚形成率和总细胞数方面无显著差异。为了找到最佳激活方案，比较了EP和Thi/DTT处理之间的卵裂率、囊胚形成率和总细胞数。Thi/DTT + 6-DMAP + DE组的囊胚形成率（36.1 ± 3.5%）和总细胞数（46.9 ± 1.0）显著高于其他组（EP + 6-DMAP + DE组、EP + Thi/DTT + 6-DMAP + DE组分别为23.3 ± 3.0%、42.2 ± 1.1和17.2 ± 2.7%、36.7 ± 1.5）。总之，本研究表明，6-DMAP和DE短期处理与标准长期处理一样有效，且Thi/DTT + 6-DMAP + DE具有协同作用。
