Disruption of NNAT, NAP1L5 and MKRN3 DNA methylation and transcription in rabbit parthenogenetic fetuses
Disruption of NNAT, NAP1L5 and MKRN3 DNA methylation and transcription in rabbit parthenogenetic fetuses
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兔孤雌胎儿中 NNAT、NAP1L5 和 MKRN3 DNA 甲基化和转录的破坏
DOI：
10.1016/j.gene.2017.05.035
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发表时间：
2017-08-30
期刊：
影响因子：
3.5
通讯作者：
Li，Zhanjun
中科院分区：
文献类型：
--
作者：
Wang，Dongxu;Liu，Zhiquan;Li，Zhanjun
孤雌激活的卵母细胞由于缺乏父本基因表达而不能在哺乳动物中发育至足月。在小鼠和猪的孤雌胚胎中，印迹基因表达和DNA甲基化状态的破坏已经被报道，但在兔子中没有。本研究采用实时荧光定量PCR（qRT-PCR）和亚硫酸氢盐测序PCR（BSP）技术，比较了孤雌生殖兔（PA）和正常受精兔（Con）的父本表达基因Neuronatin（NNAT）、核小体组装蛋白1-like 5（NAP 1 L5）和Makorin环指蛋白3（MKRN 3）的基因组印迹状态。结果显示PA组NNAT、NAP 1 L5和MKRN 3的表达明显低于Con组（p< 0.05）。此外，BSP结果表明，在兔PA胎儿中，NNAT、NAP 1 L5和MKRN 3的差异甲基化区域（DMR）存在高甲基化。综上所述，这些结果表明，DMR的高甲基化与NNAT，NAP 1 L5和MKRN 3表达的降低有关，这可能是导致兔PA胎儿发育失败的原因。
Parthenogenetically activated oocytes cannot develop to term in mammals due to lack of paternal gene expression. Disruption of imprinted gene expression and DNA methylation status in parthenogenetic fetuses has been reported in mice and pigs, but not in rabbits. In this study, the genomic imprinting status of the paternally expressed genes Neuronatin (NNAT), Nucleosome assembly protein 1-like 5 (NAP1L5), and Makorin ring finger protein 3 (MKRN3) was compared between rabbit parthenogenetic (PA) and normally fertilized fetuses (Con) using quantitative real-time PCR (qRT-PCR) and bisulfite sequencing PCR (BSP). The results revealed a significantly reduced expression ofNNAT,NAP1L5, andMKRN3in rabbit PA fetuses compared with Con fetuses (p< 0.05). In addition, the BSP results demonstrated hypermethylation in the differentially methylated regions (DMRs) ofNNAT,NAP1L5,andMKRN3in rabbit PA fetuses. Taken together, these results suggest that hypermethylation of DMRs is associated with decreasedNNAT,NAP1L5,andMKRN3expression, which may be responsible for developmental failure of rabbit PA fetuses.