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MicroRNA-34c Expression in Donor Cells Influences the Early Development of Somatic Cell Nuclear Transfer Bovine Embryos

机译:供体细胞中的MicroRNA-34c表达影响体细胞核移植牛胚胎的早期发育。

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摘要

The essence of the reprogramming activity of somatic cell nuclear transfer (SCNT) embryos is to produce normal fertilized embryos. However, reprogramming of somatic cells is not as efficient as the reprogramming of sperm. In this report, we describe the effect of an inducible, specific miR-34 microRNA expression in donor cells that enables a similar level of sperm:transgene expression on the early development of SCNT embryos. Our results showed that donor cells with doxycycline (dox)-induced miR-34c expression for the preparation of SCNT embryos resulted in altered developmental rates, histone modification (H3K9ac and H3K4me3), and extent of apoptosis. The cleavage rate and blastocyst formation of the induced nuclear transfer (NT) group were significantly increased. The immunofluorescence signal of H3K9ac in embryos in the induced NT group significantly increased in two-cell- and eight-cell-stage embryos; that of H3K4me3 increased significantly in eight-cell-stage embryos. Although significant differences in staining signals of apoptosis were not detected between groups, lower apoptosis levels were observed in the induced NT group. In conclusion, miR-34c expression induced by dox treatment enhances the developmental potential of SCNT embryos, modifies the epigenetic status, and changes blastocyst quality.
机译:体细胞核移植(SCNT)胚胎的重编程活性的实质是产生正常的受精胚胎。但是,体细胞的重编程不如精子的重编程有效。在本报告中,我们描述了可诱导的,特定的miR-34 microRNA表达在供体细胞中的作用,该表达使得SCNT胚胎早期发育具有相似水平的精子:转基因表达。我们的结果表明,用强力霉素(dox)诱导的miR-34c表达的供体细胞用于制备SCNT胚胎导致发育速度改变,组蛋白修饰(H3K9ac和H3K4me3)和凋亡程度。诱导核移植(NT)组的卵裂率和囊胚形成显着增加。诱导的NT组胚胎中H3K9ac的免疫荧光信号在两细胞和八细胞阶段的胚胎中显着增加。 H3K4me3在八细胞期胚胎中的表达显着增加。尽管在各组之间未检测到凋亡染色信号的显着差异,但在诱导的NT组中观察到较低的凋亡水平。总之,通过dox处理诱导的miR-34c表达增强了SCNT胚胎的发育潜力,改变了表观遗传状态,并改变了胚泡质量。

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