Dynamic reprogramming and function of RNA N6-methyladenosine modification during porcine early embryonic development.

N6-Methyladenosine (m6A) regulates oocyte-to-embryo transition and the reprogramming of somatic cells into induced pluripotent stem cells. However, the role of m6A methylation in porcine early embryonic development and its reprogramming characteristics in somatic cell nuclear transfer (SCNT) embryos are yet to be known. Here, we showed that m6A methylation was essential for normal early embryonic development and its aberrant reprogramming in SCNT embryos. We identified a persistent occurrence of m6A methylation in embryos between 1-cell to blastocyst stages and m6A levels abruptly increased during the morula-to-blastocyst transition. Cycloleucine (methylation inhibitor, 20 mM) treatment efficiently reduced m6A levels, significantly decreased the rates of 4-cell embryos and blastocysts, and disrupted normal lineage allocation. Moreover, cycloleucine treatment also led to higher levels in both apoptosis and autophagy in blastocysts. Furthermore, m6A levels in SCNT embryos at the 4-cell and 8-cell stages were significantly lower than that in parthenogenetic activation (PA) embryos, suggesting an abnormal reprogramming of m6A methylation in SCNT embryos. Correspondingly, expression levels of m6A writers (METTL3 and METTL14) and eraser (FTO) were apparently higher in SCNT 8-cell embryos compared with their PA counterparts. Taken together, these results indicated that aberrant nuclear transfer-mediated reprogramming of m6A methylation was involved in regulating porcine early embryonic development.

[1]  J. Li,et al.  Epigenetic Reprogramming During Somatic Cell Nuclear Transfer: Recent Progress and Future Directions , 2020, Frontiers in Genetics.

[2]  Wenjie Shu,et al.  Oocyte competence is maintained by m6A methyltransferase KIAA1429-mediated RNA metabolism during mouse follicular development , 2020, Cell Death & Differentiation.

[3]  Wenjun Zhou,et al.  Nuclear accumulation of pyruvate dehydrogenase alpha 1 promotes histone acetylation and is essential for zygotic genome activation in porcine embryos. , 2020, Biochimica et biophysica acta. Molecular cell research.

[4]  Zhenfang Wu,et al.  [Advances in epigenetic reprogramming of somatic cells nuclear transfer in mammals]. , 2019, Yi chuan = Hereditas.

[5]  Jianjun Chen,et al.  The Biogenesis and Precise Control of RNA m6A Methylation. , 2019, Trends in genetics : TIG.

[6]  Nam-Hyung Kim,et al.  Functional roles of hnRNPA2/B1 regulated by METTL3 in mammalian embryonic development , 2019, Scientific Reports.

[7]  Chuan He,et al.  Where, When, and How: Context-Dependent Functions of RNA Methylation Writers, Readers, and Erasers. , 2019, Molecular cell.

[8]  B. Tang,et al.  Tet3 is required for normal in vitro fertilization preimplantation embryos development of bovine , 2019, Molecular reproduction and development.

[9]  Youhua Liu,et al.  m6A methylation controls pluripotency of porcine induced pluripotent stem cells by targeting SOCS3/JAK2/STAT3 pathway in a YTHDF1/YTHDF2-orchestrated manner , 2019, Cell Death & Disease.

[10]  Chuan He,et al.  Chemical Modifications in the Life of an mRNA Transcript. , 2018, Annual review of genetics.

[11]  Z. Du,et al.  Reduced nucleic acid methylation impairs meiotic maturation and developmental potency of pig oocytes. , 2018, Theriogenology.

[12]  Shogo Matoba,et al.  Somatic Cell Nuclear Transfer Reprogramming: Mechanisms and Applications. , 2018, Cell stem cell.

[13]  D. Griffin,et al.  The production of pig preimplantation embryos in vitro: Current progress and future prospects. , 2018, Reproductive biology.

[14]  Alexander Kind,et al.  Genetically engineered pigs as models for human disease , 2018, Disease Models & Mechanisms.

[15]  Wei Xie,et al.  Epigenome in Early Mammalian Development: Inheritance, Reprogramming and Establishment. , 2017, Trends in cell biology.

[16]  Zuoyan Zhu,et al.  Mettl3 Mutation Disrupts Gamete Maturation and Reduces Fertility in Zebrafish , 2017, Genetics.

[17]  So Yeon Kim,et al.  Delayed blastocyst formation or an extra day culture increases apoptosis in pig blastocysts. , 2017, Animal reproduction science.

[18]  Xuerui Yang,et al.  Mettl3-/Mettl14-mediated mRNA N6-methyladenosine modulates murine spermatogenesis , 2017, Cell Research.

[19]  Yinghui Huang,et al.  m6A RNA Modification Determines Cell Fate by Regulating mRNA Degradation. , 2017, Cellular reprogramming.

[20]  H. Lee,et al.  Sirtuin inhibition leads to autophagy and apoptosis in porcine preimplantation blastocysts. , 2017, Biochemical and biophysical research communications.

[21]  Kwonho Hong,et al.  Epitranscriptome: m6A and its function in stem cell biology , 2016, Genes & Genomics.

[22]  Chuan He,et al.  m6A-dependent maternal mRNA clearance facilitates zebrafish maternal-to-zygotic transition , 2016, Nature.

[23]  S. Cánovas,et al.  Epigenetics in preimplantation mammalian development. , 2016, Theriogenology.

[24]  Qi Zhou,et al.  m(6)A RNA methylation is regulated by microRNAs and promotes reprogramming to pluripotency. , 2015, Cell stem cell.

[25]  R. Prather,et al.  Impairment of Preimplantation Porcine Embryo Development by Histone Demethylase KDM5B Knockdown Through Disturbance of Bivalent H3K4me3-H3K27me3 Modifications1 , 2015, Biology of reproduction.

[26]  Jennifer Hamm,et al.  Dynamics of TET family expression in porcine preimplantation embryos is related to zygotic genome activation and required for the maintenance of NANOG. , 2014, Developmental biology.

[27]  Yang Wang,et al.  N6-methyladenosine modification destabilizes developmental regulators in embryonic stem cells , 2014, Nature Cell Biology.

[28]  I. Choi,et al.  Transcription factor AP-2γ is a core regulator of tight junction biogenesis and cavity formation during mouse early embryogenesis , 2012, Development.

[29]  Nam-Hyung Kim,et al.  Modulation of autophagy influences development and apoptosis in mouse embryos developing in vitro , 2011, Molecular reproduction and development.

[30]  R. Roberts,et al.  A commentary on domestic animals as dual-purpose models that benefit agricultural and biomedical research. , 2008, Journal of animal science.

[31]  T. Haaf,et al.  Aberrant methylation patterns at the two‐cell stage as an indicator of early developmental failure , 2002, Molecular reproduction and development.

[32]  Eszter Posfai,et al.  The mammalian embryo's first agenda: making trophectoderm. , 2019, The International journal of developmental biology.

[33]  J. Nichols,et al.  States and Origins of Mammalian Embryonic Pluripotency In Vivo and in a Dish. , 2018, Current topics in developmental biology.

[34]  V. Yartseva,et al.  The Maternal-to-Zygotic Transition During Vertebrate Development: A Model for Reprogramming. , 2015, Current topics in developmental biology.