Lee Joohyeong, Park Jong-Im, Yun Jung Im, Lee Yongjin, Yong Hwanyul, Lee Seung Tae, Park Choon-Keun, Hyun Sang-Hwan, Lee Geun-Shik, Lee Eunsong
College of Veterinary Medicine, Kangwon National University, Chuncheon 200-701, Korea.
J Vet Sci. 2015;16(3):373-80. doi: 10.4142/jvs.2015.16.3.373. Epub 2015 Mar 20.
This study was conducted to investigate the effects of rapamycin treatment during in vitro maturation (IVM) on oocyte maturation and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) in pigs. Morphologically good (MGCOCs) and poor oocytes (MPCOCs) were untreated or treated with 1 nM rapamycin during 0-22 h, 22-42 h, or 0-42 h of IVM. Rapamycin had no significant effects on nuclear maturation and blastocyst formation after PA of MGCOCs. Blastocyst formation after PA was significantly increased by rapamycin treatment during 22-42 h and 0-42 h (46.6% and 46.5%, respectively) relative to the control (33.3%) and 0-22 h groups (38.6%) in MPCOCs. In SCNT, blastocyst formation tended to increase in MPCOCs treated with rapamycin during 0-42 h of IVM relative to untreated oocytes (20.3% vs. 14.3%, 0.05 < p < 0.1), while no improvement was observed in MGCOCs. Gene expression analysis revealed that transcript abundance of Beclin 1 and microtubule-associated protein 1 light chain 3 mRNAs was significantly increased in MPCOCs by rapamycin relative to the control. Our results demonstrated that autophagy induction by rapamycin during IVM improved developmental competence of oocytes derived from MPCOCs.
本研究旨在探讨雷帕霉素处理对猪体外成熟(IVM)期间卵母细胞成熟以及孤雌激活(PA)和体细胞核移植(SCNT)后胚胎发育的影响。形态良好的卵丘-卵母细胞复合体(MGCOCs)和形态较差的卵母细胞(MPCOCs)在IVM的0 - 22小时、22 - 42小时或0 - 42小时期间不进行处理或用1 nM雷帕霉素处理。雷帕霉素对MGCOCs孤雌激活后的核成熟和囊胚形成没有显著影响。与MPCOCs的对照组(33.3%)和0 - 22小时组(38.6%)相比,在22 - 42小时和0 - 42小时期间用雷帕霉素处理后,孤雌激活后的囊胚形成显著增加(分别为46.6%和46.5%)。在体细胞核移植中,与未处理的卵母细胞相比,IVM的0 - 42小时期间用雷帕霉素处理的MPCOCs囊胚形成有增加趋势(20.3%对14.3%,0.05 < p < 0.1),而MGCOCs未观察到改善。基因表达分析显示,与对照组相比,雷帕霉素使MPCOCs中Beclin 1和微管相关蛋白1轻链3 mRNA的转录丰度显著增加。我们的结果表明,IVM期间雷帕霉素诱导自噬可提高源自MPCOCs的卵母细胞的发育能力。