The perturbed expression of m6A in parthenogenetic mouse embryos.

Jindong Hao,Liang Han,Minghui Qi,Shuming Shi,Jiaqi Wei,Yu Xianfeng,Dongxu Wang,Wei Gao,Chao Lin

doi:10.1590/1678-4685-gmb-2018-0212

Jindong Hao, Liang Han + Show 7 more

Open Access

https://doi.org/10.1590/1678-4685-gmb-2018-0212

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Abstract

Parthenogenetically activated oocytes cannot develop to term in mammals owing to abnormal epigenetic modifications. Methylation of the N6 position of adenosine (m6A) is a post-transcriptional epigenetic modification of RNA. To investigate the role of m6A methylation in parthenogenetic (PA) embryonic development, we analyzed METTL3, METTL14, FTO, ALKBH5, YTHDF2, IGF2BP1, and IGF2BP2 expression by quantitative real-time PCR. These genes were found dynamically expressed during the 2-cell, 4-cell, 8-cell, and blastocyst stages of the embryo. Compared to normally fertilized embryos, the expression of these genes was perturbed in PA embryos, especially at the 8-cell stage. Furthermore, immunofluorescence was used to detect m6A expression. The results demonstrated that m6A expression decreased in the 2-cell stage, whereas it increased in the 8-cell stage of PA embryos. Taken together, these results suggest that the expression of RNA methylation-related genes was perturbed, leading to abnormal m6A modification during early development in PA embryos.

Highlights

Parthenogenetic (PA) embryos contain exclusively maternal genomes and have been used extensively to study epigenetic profiles and determine the expression patterns of imprinted genes in human diseases (Park et al, 2009; Zhang et al, 2015)
The expression patterns of METTL3, METTL14, FTO, ALKBH5, YTHDF2, IGF2BP1, and IGF2BP2 were determined by Quantitative real-time PCR (qRT-PCR) during early embryonic development
Parthenogenetic activation was confirmed by the presence of two pronuclei, which developed to the 2-cell stage

Summary

Introduction

Parthenogenetic (PA) embryos contain exclusively maternal genomes and have been used extensively to study epigenetic profiles and determine the expression patterns of imprinted genes in human diseases (Park et al, 2009; Zhang et al, 2015). The expression patterns of METTL3, METTL14, FTO, ALKBH5, YTHDF2, IGF2BP1, and IGF2BP2 were determined by qRT-PCR during early embryonic development. Quantitative real-time PCR (qRT-PCR) was performed to determine METTL3, METTL14, FTO, ALKBH5, YTHDF2 IGF2BP1, and IGF2BP2 expression using the BioEasy SYBR Green I Real-Time PCR Kit on the BIO-RAD iQ5 Multicolor Real-Time PCR Detection System (Bioer Technology, China).

Results

Conclusion