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Expression and DNA Methylation Change of Oct-4 in Cloned Bovine Blastocysts  

Cha, Byung-Hyun (National Institute of Animal Science, RDA)
Choi, Jung-Sang (National Institute of Animal Science, RDA)
Hwang, Seong-Soo (National Institute of Animal Science, RDA)
Chung, Hak-Jae (National Institute of Animal Science, RDA)
Im, Gi-Sun (National Institute of Animal Science, RDA)
Yang, Byong-Chul (National Institute of Animal Science, RDA)
Kim, Myong-Jik (National Institute of Animal Science, RDA)
Cho, Jae-Hyeon (Dept. of Veterinary Medicine, Gyeongsang National University)
Seong, Hwan-Hoo (National Institute of Animal Science, RDA)
Ko, Yeoung-Gyu (National Institute of Animal Science, RDA)
Publication Information
Journal of Embryo Transfer / v.23, no.3, 2008 , pp. 133-139 More about this Journal
Abstract
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.
Keywords
bovine; Oct-4; DNA methylation; somatic cell nuclear transfer; blastocyst;
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