Lee Joohyeong, You Jinyoung, Lee Geun-Shik, Lee Seung Tae, Hyun Sang-Hwan, Lee Eunsong
a College of Veterinary Medicine , Kangwon National University , Chuncheon , Korea.
b Institute of Veterinary Science , Kangwon National University , Chuncheon , Korea.
Anim Biotechnol. 2018 Jan 2;29(1):41-49. doi: 10.1080/10495398.2017.1294598. Epub 2017 Mar 30.
This study determined the effects of postactivation treatment with demecolcine and/or 6-dimethylaminopurine (6-DMAP) on in vivo and in vitro developmental competence of somatic cell nuclear transfer (SCNT) embryos in pigs. SCNT embryos were treated for 4 hours with 0.4 µg/mL demecolcine, 2 mM 6-DMAP, or both after electric activation, then transferred to surrogate pigs or cultured for 7 days. The formation rate of SCNT embryos with a single pronucleus was higher in combined treatment with demecolcine and 6-DMAP (95.2%) than treatment with demecolcine alone (87.1%). Blastocyst formation of SCNT embryos was significantly increased in combined treatment with demecolcine and 6-DMAP (48.7%) compared with demecolcine (22.2%) or 6-DMAP alone (37.3%). Fluctuation of maturation promoting factor activity showed different patterns among various postactivation treatments. Pregnancy was established in 1 of 5 surrogates after transfer of SCNT embryos that were treated with demecolcine and 6-DMAP. The pregnant surrogate delivered one healthy live piglet. The results of our study demonstrated that postactivation treatment with demecolcine and 6-DMAP together improved preimplantation development and supported normal in vivo development of SCNT pig embryos, probably influencing MPF activity and nuclear remodeling, including induction of single pronucleus formation after electric activation.
本研究确定了秋水仙碱和/或6-二甲基氨基嘌呤（6-DMAP）激活后处理对猪体细胞核移植（SCNT）胚胎体内和体外发育能力的影响。电激活后，将SCNT胚胎用0.4µg/mL秋水仙碱、2 mM 6-DMAP或两者联合处理4小时，然后移植到代孕猪体内或培养7天。秋水仙碱和6-DMAP联合处理的SCNT胚胎单原核形成率（95.2%）高于单独使用秋水仙碱处理（87.1%）。与单独使用秋水仙碱（22.2%）或6-DMAP（37.3%）相比，秋水仙碱和6-DMAP联合处理显著提高了SCNT胚胎的囊胚形成率（48.7%）。在各种激活后处理中，成熟促进因子活性的波动呈现出不同模式。用秋水仙碱和6-DMAP处理的SCNT胚胎移植后，5头代孕猪中有1头成功怀孕。怀孕的代孕猪产下了一头健康的活仔猪。我们的研究结果表明，秋水仙碱和6-DMAP联合激活后处理改善了植入前发育，并支持了SCNT猪胚胎的正常体内发育，可能影响了MPF活性和核重塑，包括电激活后诱导单原核形成。
