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KMID : 1005220080230030133
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Journal of Embryo Transfer
2008 Volume.23 No. 3 p.133 ~ p.139
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Expression and DNA Methylation Change of Oct-4 in Cloned Bovine Blastocysts
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Cha Byung-Hyun
Choi Jung-Sang
Hwang Seong-Soo
Chung Hak-Jae
Im Gi-Sun
Yang Byong-Chul
Kim Myong-Jik
Cho Jae-Hyeon
Seong Hwan-Hoo
Ko Yeoung-Gyu
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Abstract
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DNA methylation is one of the reasons for poor survival of clone animals. The OCT-4 gene is essential for maintaining pluripotency of embryonic stem (ES) cells and early embryos. We previously reported that the 5’-promoter region of Oct-4 gene was a target of DNA methylation and the methylation status was changed variously during embryonic development in bovine. The study conducted to examine the expression and methylation pattern of tissue-dependent differentially methylated region (T-DMR) of Oct-4 gene in bovine somatic cell nuclear transfer (SCNT) and in vitro fertilization (IVF) blastocysts. The Oct-4 gene expression was evaluated by RT-PCR and fluorescence immunocytochemistry. The methylation pattern of T-DMR was analyzed using restriction mapping and bisulfite sequencing methods. The Oct-4 transcripts were highly expressed in IVF, while they were not expressed in SCNT. The Oct-4 protein was not detected or expressed at very low level in SCNT, the intensity of Oct-4 protein, however, was strong in IVF. On the other hand, the T-DMR of Oct-4 gene was hypermethylated in SCNT compared to that of IVF. These results suggested that expression and the failure of demethylation of Oct-4 gene was closely associated with incomplete development of SCNT embryos.
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KEYWORD
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bovine, Oct-4, DNA methylation, somatic cell nuclear transfer, blastocyst
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