Diao Yun Fei, Lin Tao, Li Xiaoxia, Oqani Reza K, Lee Jae Eun, Kim So Yeon, Jin Dong Il
Institute of Special Animal & Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China.
Department of Animal Science & Biotechnology, Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon, Republic of Korea.
PLoS One. 2018 Feb 15;13(2):e0191816. doi: 10.1371/journal.pone.0191816. eCollection 2018.
SETD2 (SET domain containing protein 2) acts as a histone H3 lysine 36 (H3K36)-specific methyltransferase and may play important roles in active gene transcription in human cells. However, its expression and role in porcine oocytes and preimplantation embryos are not well understood. Here, we used immunofluorescence and laser scanning confocal microscopy to examine SETD2 expression in porcine fetal fibroblasts, oocytes, and preimplantation embryos derived from in vitro fertilization (IVF), parthenogenetic activation (PA), and somatic cell nuclear transfer (SCNT). In porcine fetal fibroblasts, SETD2 expression was detected in interphase cells, but not in M (mitotic)-phase cells. The SETD2 signal was observed in non-surrounded nucleolus (NSN)-stage oocytes, but not in surrounded nucleolus (SN)-, metaphase I (MI)-, or metaphase II (MII)-stage oocytes. The SETD2 signal was detectable in sperm, and undetectable immediately after fertilization, detectable at the 2-cell stage, and peaked at the 4-cell stage of IVF embryos in which porcine embryonic genome is activated. Similar to the pattern found in IVF embryos, the SETD2 signal was absent from PA embryos at the 1-cell stage, but it was detected at the 2-cell stage and thereafter maintained to the blastocyst stage. Interestingly, unlike the IVF and PA embryos, the SETD2 signal was detected throughout the development of SCNT embryos, including at the 1-cell stage. These data suggest that SETD2 may be functional for embryonic gene transcription in porcine preimplantation embryos. It is further speculated that the aberrant expression of SETD2 at the 1-cell stage of porcine SCNT embryos may be a factor in the low efficiency of cloning in pig.
SETD2(含SET结构域蛋白2)作为一种组蛋白H3赖氨酸36(H3K36)特异性甲基转移酶,可能在人类细胞的活性基因转录中发挥重要作用。然而,其在猪卵母细胞和植入前胚胎中的表达及作用尚不清楚。在此,我们利用免疫荧光和激光扫描共聚焦显微镜检测了SETD2在猪胎儿成纤维细胞、卵母细胞以及体外受精(IVF)、孤雌激活(PA)和体细胞核移植(SCNT)来源的植入前胚胎中的表达。在猪胎儿成纤维细胞中,SETD2表达在间期细胞中可检测到,但在M(有丝分裂)期细胞中未检测到。在非包围核仁(NSN)期卵母细胞中观察到SETD2信号,但在包围核仁(SN)期、减数第一次分裂中期(MI)或减数第二次分裂中期(MII)期卵母细胞中未观察到。SETD2信号在精子中可检测到,受精后立即不可检测,在2细胞期可检测到,并在猪胚胎基因组被激活的IVF胚胎的4细胞期达到峰值。与IVF胚胎中的模式相似,PA胚胎在1细胞期不存在SETD2信号,但在2细胞期可检测到,此后一直维持到囊胚期。有趣的是,与IVF和PA胚胎不同,SETD2信号在SCNT胚胎的整个发育过程中均可检测到,包括在1细胞期。这些数据表明,SETD2可能在猪植入前胚胎的胚胎基因转录中发挥作用。进一步推测,猪SCNT胚胎1细胞期SETD2的异常表达可能是猪克隆效率低下的一个因素。