Division of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri 65211, USA.
Mol Reprod Dev. 2012 Mar;79(3):197-207. doi: 10.1002/mrd.22013. Epub 2011 Dec 27.
Evidence in many species has suggested that those embryos that cleave earliest after fertilization are more developmentally competent than those that cleave relatively later after fertilization. Herein we document this phenomenon in porcine in vitro-fertilized (IVF), somatic cell nuclear transfer (SCNT), and parthenogenetic (PA) embryos. In vitro-matured pig oocytes were used to generate IVF, SCNT, and PA embryos. At 24 hr post-activation (or insemination; hpa/hpi), embryos were visually assessed, and cleaved embryos were moved into a new culture well. This process was repeated at 30 and 48 hpa/hpi. All embryos were allowed to develop 7 days in culture. For IVF embryos, 39.9%, 24.6%, and 10.5% of fast-, intermediate-, or slow-cleaving embryos, respectively, developed into blastocysts by day 7. For SCNT embryos, 31.8% of fast-, 5.7% of intermediate-, and 2.9% of late-cleaving embryos achieved the blastocyst stage of development. For PA embryos, the percentages of those cleaved embryos that developed to blastocyst were 59.3%, 36.7%, and 7.5% for early-, intermediate-, and late-cleaving embryos, respectively. Using RNA collected from early-, intermediate-, and late-cleaving embryos, real-time PCR was performed to assess the transcript levels of 14 different genes of widely varied function. The qPCR results suggest that maternal mRNA degradation may not proceed in an appropriate pattern in slow-cleaving embryos. These findings (1) confirm that, as observed in other species, earlier-cleaving porcine embryos are more successful at developing in culture than are slower-cleaving embryos, and (2) implicate mechanisms of maternal transcript destruction as potential determinants of oocyte/embryo quality.
在许多物种中都有证据表明,那些在受精后最早分裂的胚胎比那些相对较晚分裂的胚胎在发育上更具竞争力。本文记录了猪体外受精(IVF)、体细胞核移植(SCNT)和孤雌激活(PA)胚胎中的这一现象。使用体外成熟的猪卵母细胞来产生 IVF、SCNT 和 PA 胚胎。在激活(或受精)后 24 小时(hpa/hpi),通过肉眼评估胚胎,并将已分裂的胚胎转移到新的培养孔中。在 30 和 48 hpa/hpi 时重复此过程。所有胚胎都在培养中发育 7 天。对于 IVF 胚胎,快速、中间或慢速分裂的胚胎分别有 39.9%、24.6%和 10.5%在第 7 天发育成囊胚。对于 SCNT 胚胎,快速分裂的胚胎有 31.8%、中间分裂的胚胎有 5.7%、晚期分裂的胚胎有 2.9%发育成囊胚。对于 PA 胚胎,早期、中间和晚期分裂的胚胎发育成囊胚的比例分别为 59.3%、36.7%和 7.5%。使用从早期、中期和晚期分裂的胚胎中收集的 RNA,进行实时 PCR 以评估 14 种不同功能的基因的转录水平。qPCR 结果表明,慢速分裂胚胎中的母体 mRNA 降解可能没有以适当的模式进行。这些发现:1)证实了在其他物种中观察到的现象,即在培养中,早期分裂的猪胚胎比晚期分裂的胚胎更成功,2)暗示了母体转录物破坏的机制可能是卵母细胞/胚胎质量的潜在决定因素。