Graduate School of Biology-Oriented Science and Technology, Kindai University, Wakayama 649-6493, Japan.
MRC Clinical Sciences Centre, Imperial College Faculty of Medicine, W12 0NN London, UK.
J Reprod Dev. 2020 Jun 12;66(3):255-263. doi: 10.1262/jrd.2019-167. Epub 2020 Mar 26.
The large Japanese field mouse (Apodemus speciosus) is endemic to Japan and may be used as an animal model for studies related to environmental pollution, medical science, and basic biology. However, the large Japanese field mouse has low reproductive ability due to the small number of oocytes ovulated per female. To produce experimental models, we investigated the in vitro developmental potential of interspecies somatic cell nuclear transfer (iSCNT) embryos produced by fusing tail tip cells from the large Japanese field mouse with enucleated oocytes from laboratory mice (Mus musculus domesticus). Only a small number of iSCNT embryos developed to the 4-cell (0-4%) and blastocysts (0-1%) stages under sequential treatment using trichostatin A (TSA) and vitamin C (VC) supplemented with deionized bovine serum albumin (d-BSA). This sequential treatment led to the reduction in H3K9 trimethylation and did not affect H3K4 trimethylation in at least the 2-cell stage of the iSCNT embryos. Moreover, iSCNT embryos that received tail tip cells with exposure treatment to ooplasm from cell fusion to oocyte activation or VC treatment prior to cell fusion did not exhibit significant in vitro development improvement compared to that of each control group. This suggests that large Japanese field mice/laboratory mice iSCNT embryos that received sequential treatment using TSA and VC with d-BSA may have slightly better developmental potential beyond the 4-cell stage. Our results provide insights into the reprogramming barriers impeding the wider implementation of iSCNT technology.
大型日本田鼠(Apodemus speciosus)是日本特有的一种物种,可能被用作与环境污染、医学科学和基础生物学相关的研究的动物模型。然而,由于雌性个体排卵的卵母细胞数量较少,大型日本田鼠的繁殖能力较低。为了生产实验模型,我们研究了通过融合大型日本田鼠尾尖细胞和去核的实验室小鼠(Mus musculus domesticus)卵母细胞产生的种间体细胞核移植(iSCNT)胚胎的体外发育潜力。只有少数 iSCNT 胚胎在使用曲古抑菌素 A(TSA)和维生素 C(VC)处理,并添加去离子牛血清白蛋白(d-BSA)的情况下,能够发育到 4 细胞(0-4%)和囊胚(0-1%)阶段。这种序贯处理导致 H3K9 三甲基化减少,但至少在 iSCNT 胚胎的 2 细胞阶段,不影响 H3K4 三甲基化。此外,与每个对照组相比,接受尾尖细胞暴露处理的 iSCNT 胚胎(从细胞融合到卵母细胞激活,或在细胞融合前接受 VC 处理),其体外发育并没有显著改善。这表明,使用 TSA 和 VC 与 d-BSA 进行序贯处理的大型日本田鼠/实验室小鼠 iSCNT 胚胎,可能在 4 细胞阶段之后具有稍好的发育潜力。我们的研究结果为影响 iSCNT 技术广泛应用的重编程障碍提供了新的见解。