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KMID : 1023620120360010007
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Reproductive and Developmental Biology
2012 Volume.36 No. 1 p.7 ~ p.12
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Dynamic DNA Methylation Change of Dnmt1o 5’-Terminal Region during Preimplantation Development of Cloned Pig
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Ko Yeoung-Gyu
Kim Sung-Woo
Cho Sang-Rae
Do Yoon-Jung
Kim Jae-Hwan
Kim Sang-Woo
Kim Hyun
Park Jae-Hong
Park Soo-Bong
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Abstract
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DNA methyltransferase 1 (Dnmt1) gene contains three different isoform transcripts, Dnmt1s, Dnmt1o, and Dnmt1p,
are produced by alternative usage of multiple first exons. Dnmt1o is specific to oocytes and preimplantation embryos,
whereas Dnmt1s is expressed in somatic cells. Here we determined that porcine Dnmt1o gene had differentially methylated regions (DMRs) in 5’-flanking region, while those were not found in the Dnmt1s promoter region. The
methylation patterns of the porcine Dnmt1o/Dnmt1s DMRs were investigated using bisulfite sequencing and pyrosequencing analysis through all preimplantation stages from one cell to blastocyst stage in in vivo or somatic cell nuclear transfer (SCNT). The Dnmt1o DMRs contained 8 CpG sites, which located in ?640 bp to ?30 bp upstream region from transcription start site of the Dnmt1o gene. The methylation status of 5 CpGs within the Dnmt1o DMRs were distinctively different at each stage from one-cell to blastocyst stage in the in vivo or SCNT, respectively. 55.62% methylation degree of the Dnmt1o DMRs in the in vivo was increased up to 84.38% in the SCNT embryo, moreover, de novo methylation and demethylation occurred during development of porcine embryos from the one-cell stage to the blastocyst stage. However, the DNA methylation states at CpG sites in the Dnmt1s promoter regions were hypomethylated,
and dramatically not changed through one-cell to blastocyst stage in the in vivo or SCNT embryos. In the present
study, we demonstrated that the DMRs in the promoter region of the porcine Dnmt1o was well conserved, contributing
to establishment and maintenance of genome-wide patterns of DNA methylation in early embryonic development.
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KEYWORD
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Porcine DNA methyltransferase 1o/1s, DNA methylation, Differentially methylated region (DMR), Somatic cell nuclear transfer (SCNT)
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