Analysis of nuclear reprogramming in cloned miniature pig embryos by expression of Oct-4 and Oct-4 related genes |
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Authors: | Lee Eugine Lee So Hyun Kim Sue Jeong Yeon Woo Kim Ji Hye Koo Ok Jae Park Seon Mi Hashem M A Hossein M S Son Hye Young Lee Chang Kyu Hwang Woo Suk Kang Sung Keun Lee Byeong Chun |
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Affiliation: | Department of Theriogenology and Biotechnololgy, College of Veterinary Medicine, Seoul National University, Seoul 151-742, South Korea. |
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Abstract: | Xenotransplantation is a rapidly expanding field of research and cloned miniature pigs have been considered as a model animal for it. However, the efficiency of somatic cell nuclear transfer (SCNT) is extremely low, with most clones resulting in early lethality and several kinds of aberrant development. A possible explanation for the developmental failure of SCNT embryos is insufficient reprogramming of the somatic cell nucleus by the oocyte. In order to test this, we analyzed the reprogramming capacity of differentiated fibroblast cell nuclei and embryonic germ cell nuclei with Oct-4 and Oct-4 related genes (Ndp5211, Dppa2, Dppa3, and Dppa5), which are important for embryonic development, Hand1 and GATA-4, which are important for placental development, as molecular markers using RT-PCR. The Oct-4 expression level was significantly lower (P<0.05) in cloned hatched blastocysts derived from fibroblasts and many of fibroblast-derived clones failed to reactivate at least one of the tested genes, while most of the germ cell clones and control embryos correctly expressed these genes. In conclusion, our results suggest that the reprogramming of fibroblast-derived cloned embryos is highly aberrant and this improper reprogramming could be one reason of the early lethality and post-implantation anomalies of somatic cell-derived clones. |
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Keywords: | Cloned pig embryos Miniature pig Nuclear transfer Oct-4 Reprogramming Embryonic germ cell |
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