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以孤雌生殖体细胞作为核供体的猪克隆胎儿中Igf2/H19的基因组印记分析

Genomic imprinting analysis of Igf2/H19 in porcine cloned fetuses using parthenogenetic somatic cells as nuclear donors.

作者信息

Wang Dongxu, Song Yuning, Huang Yongye, Duan Feifei, Lv Qinyan, Ouyang Hongsheng, Lai Liangxue, Li Zhanjun

机构信息

Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Science, Jilin University, 5333#, Xi'an Road, Changchun, 130062, China,

出版信息

Biotechnol Lett. 2014 Oct;36(10):1945-52. doi: 10.1007/s10529-014-1572-8. Epub 2014 Jun 15.

Abstract

To gain insight into parthenogenesis in pigs, we report for the first time that using parthenogenetic somatic cells as nuclear donors (PSCNT), the porcine parthenogenetic fetus can develop to gestational day 39. Weight and morphological analysis revealed that PSCNT fetuses were smaller and developmentally retarded when compared to normally fertilized controls. Quantitative gene expression analysis indicated that in PSCNT fetuses, H19 was over-expressed, whereas Igf2 was significantly reduced (p < 0.05) compared with their controls. In addition, bisulfite-sequencing PCR results demonstrated that H19 differentially DNA methylated regions (DMRs) were hypomethylated in PSCNT fetuses, while Igf2 DMRs were hypermethylated in both PSCNT and control fetuses. Our results suggest that extended development of the porcine parthenogenetic fetus can be accomplished using PSCNT and that abnormal DNA methylation of H19 DMRs might contribute to the critical barrier of parthenogenesis in pigs.

摘要

为深入了解猪的孤雌生殖,我们首次报道,使用孤雌生殖体细胞作为核供体(PSCNT),猪孤雌生殖胎儿可发育至妊娠第39天。体重和形态分析显示,与正常受精的对照相比,PSCNT胎儿更小且发育迟缓。定量基因表达分析表明,在PSCNT胎儿中,H19过度表达,而与对照相比,Igf2显著降低(p < 0.05)。此外,亚硫酸氢盐测序PCR结果表明,PSCNT胎儿中H19差异DNA甲基化区域(DMRs)低甲基化,而在PSCNT和对照胎儿中Igf2 DMRs均高甲基化。我们的结果表明,使用PSCNT可实现猪孤雌生殖胎儿的延长发育,且H19 DMRs的异常DNA甲基化可能是猪孤雌生殖关键障碍的原因。

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