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Sperm-borne small RNAs improve the developmental competence of pre-implantation cloned embryos in rabbit
Zygote ( IF 1.7 ) Pub Date : 2021-03-09 , DOI: 10.1017/s0967199420000805
Hongyu Qin 1 , Pengxiang Qu 2 , Huizhong Hu 2 , Wenbin Cao 2 , Hengchao Liu 2 , Yanru Zhang 2 , Jinpeng Zhao 2 , Fatima Nazira 2 , Enqi Liu 2
Affiliation  

SummaryThe low efficiency of somatic cell nuclear transfer (SCNT) greatly limits its application. Compared with the fertilized embryo, cloned embryos display abnormal epigenetic modification and other inferior developmental properties. In this study, small RNAs were isolated, and miR-34c and miR-125b were quantified by real-time PCR; results showed that these micro-RNAs were highly expressed in sperm. The test sample was divided into three groups: one was the fertilized group, one was the SCNT control group (NT-C group), and the third group consisted of SCNT embryos injected with sperm-borne small RNA (NT-T group). The level of tri-methylation of lysine 9 on histone H3 (H3K9me3) at the 8-cell stage was determined by immunofluorescence staining, and the cleavage ratio, blastocyst ratio, apoptotic cell index of the blastocyst and total cell number of blastocysts in each group were analyzed. Results showed that the H3K9me3 level was significantly higher in the NT-C group than in the fertilized group and the NT-T group. The apoptosis index of blastocysts in the NT-C group was significantly higher than that in the fertilized group and the NT-T group. The total cell number of SCNT embryos was significantly lower than that of fertilized embryos, and injecting sperm-borne small RNAs could significantly increase the total cell number of SCNT blastocysts. Our study not only demonstrates that sperm-borne small RNAs have an important role in embryo development, but also provides a new strategy for improving the efficiency of SCNT in rabbit.

中文翻译:

精子携带小RNAs提高兔植入前克隆胚胎的发育能力

总结体细胞核移植(SCNT)的低效率极大地限制了它的应用。与受精胚胎相比,克隆胚胎表现出异常的表观遗传修饰和其他较差的发育特性。在本研究中,分离小RNA,并通过实时PCR定量miR-34c和miR-125b;结果表明,这些微RNA在精子中高度表达。测试样本分为三组:一组为受精组,一组为SCNT对照组(NT-C组),第三组为注射了精子携带的小RNA的SCNT胚胎(NT-T组)。免疫荧光染色测定8细胞期组蛋白H3(H3K9me3)赖氨酸9的三甲基化水平,卵裂率、囊胚率、分析各组胚泡凋亡细胞指数和胚泡总细胞数。结果显示,NT-C组H3K9me3水平明显高于受精组和NT-T组。NT-C组囊胚凋亡指数明显高于受精组和NT-T组。SCNT胚胎的总细胞数显着低于受精胚胎,注射精子携带的小RNA可显着增加SCNT囊胚的总细胞数。我们的研究不仅证明了精子携带的小RNA在胚胎发育中具有重要作用,而且为提高兔SCNT的效率提供了新的策略。结果显示,NT-C组H3K9me3水平明显高于受精组和NT-T组。NT-C组囊胚凋亡指数明显高于受精组和NT-T组。SCNT胚胎的总细胞数显着低于受精胚胎,注射精子携带的小RNA可显着增加SCNT囊胚的总细胞数。我们的研究不仅证明了精子携带的小RNA在胚胎发育中具有重要作用,而且为提高兔SCNT的效率提供了新的策略。结果显示,NT-C组H3K9me3水平明显高于受精组和NT-T组。NT-C组囊胚凋亡指数明显高于受精组和NT-T组。SCNT胚胎的总细胞数显着低于受精胚胎,注射精子携带的小RNA可显着增加SCNT囊胚的总细胞数。我们的研究不仅证明了精子携带的小RNA在胚胎发育中具有重要作用,而且为提高兔SCNT的效率提供了新的策略。SCNT胚胎的总细胞数显着低于受精胚胎,注射精子携带的小RNA可显着增加SCNT囊胚的总细胞数。我们的研究不仅证明了精子携带的小RNA在胚胎发育中具有重要作用,而且为提高兔SCNT的效率提供了新的策略。SCNT胚胎的总细胞数显着低于受精胚胎,注射精子携带的小RNA可显着增加SCNT囊胚的总细胞数。我们的研究不仅证明了精子携带的小RNA在胚胎发育中具有重要作用,而且为提高兔SCNT的效率提供了新的策略。
更新日期:2021-03-09
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