Enucleated oocytes can reprogram differentiated nuclei to totipotency after somatic cell nuclear transfer (SCNT), which is valuable in understanding nuclear reprogramming and generating genetically modified animals. To date, reprogramming efficiency is low and the development of SCNT embryos is not going as well as anticipated. To further disclose the reprogramming mechanisms during SCNT zebrafish embryo development, we examined the expression patterns of transcription regulation factors and regulated them by mRNA and morpholino microinjection. In this study, we show that stem cell-related transcription factors are downregulated in zebrafish SCNT embryos at the blastula stage. Exogenous expression of pou5f3 at the single-cell stage improves SCNT embryo development from the blastula to the gastrula stage. We also found that exogenous expression of klf4 or sox2 decreases SCNT embryo development from the blastula to the gastrula stage, while expression of nanog is necessary for the development of SCNT embryos. Our results conclude that zebrafish pou5f3 facilitates the development of SCNT embryos from the blastula to gastrula stage.

中文翻译：

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pou5f3的外源表达促进斑马鱼早期体细胞核移植胚胎的发育

去核卵母细胞可以在体细胞核移植 (SCNT) 后将分化的细胞核重编程为全能性，这对于理解核重编程和产生转基因动物很有价值。迄今为止，重编程效率很低，SCNT 胚胎的发育也没有预期的那么好。为了进一步揭示 SCNT 斑马鱼胚胎发育过程中的重编程机制，我们检测了转录调控因子的表达模式，并通过 mRNA 和 morpholino 显微注射对其进行调控。在这项研究中，我们表明干细胞相关转录因子在囊胚期斑马鱼 SCNT 胚胎中被下调。pou5f3的外源表达在单细胞阶段改善了从囊胚到原肠胚阶段的 SCNT 胚胎发育。我们还发现，klf4或sox2的外源表达会降低 SCNT 胚胎从囊胚到原肠胚阶段的发育，而 nanog 的表达是SCNT 胚胎发育所必需的。我们的研究结果得出结论，斑马鱼pou5f3促进了 SCNT 胚胎从囊胚到原肠胚阶段的发育。