Spate Lee D, Brown Alana N, Redel Bethany K, Whitworth Kristin M, Murphy Clifton N, Prather Randall S
Division of Animal Science, Animal Science Research Center, Columbia, Missouri, United States of America.
PLoS One. 2014 Apr 16;9(4):e95114. doi: 10.1371/journal.pone.0095114. eCollection 2014.
The ability to mature oocytes in vitro provides a tool for creating embryos by parthenogenesis, fertilization, and cloning. Unfortunately the quality of oocytes matured in vitro falls behind that of in vivo matured oocytes. To address this difference, transcriptional profiling by deep sequencing was conducted on pig oocytes that were either matured in vitro or in vivo. Alignment of over 18 million reads identified 1,316 transcripts that were differentially represented. One pathway that was overrepresented in the oocytes matured in vitro was for Wingless-type MMTV integration site (WNT) signaling. In an attempt to inhibit the WNT pathway, Dickkopf-related protein 1 was added to the in vitro maturation medium. Addition of Dickkopf-related protein 1 improved the percentage of oocytes that matured to the metaphase II stage, increased the number of nuclei in the resulting blastocyst stage embryos, and reduced the amount of disheveled segment polarity protein 1 protein in oocytes. It is concluded that transcriptional profiling is a powerful method for detecting differences between in vitro and in vivo matured oocytes, and that the WNT signaling pathway is important for proper oocyte maturation.
体外成熟卵母细胞的能力为通过孤雌生殖、受精和克隆来创建胚胎提供了一种工具。不幸的是,体外成熟的卵母细胞质量落后于体内成熟的卵母细胞。为了解决这种差异,对体外或体内成熟的猪卵母细胞进行了深度测序转录谱分析。超过1800万条读数的比对鉴定出1316个差异表达的转录本。在体外成熟的卵母细胞中过度表达的一条信号通路是无翅型MMTV整合位点(WNT)信号通路。为了抑制WNT信号通路,将Dickkopf相关蛋白1添加到体外成熟培养基中。添加Dickkopf相关蛋白1提高了成熟到中期II期的卵母细胞百分比,增加了所得囊胚期胚胎中的细胞核数量,并减少了卵母细胞中无序节段极性蛋白1的量。得出的结论是,转录谱分析是检测体外和体内成熟卵母细胞之间差异的有力方法,并且WNT信号通路对卵母细胞的正常成熟很重要。