Analysis of nuclear reprogramming in cloned miniature pig embryos by expression of Oct-4 and Oct-4 related genes.
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.
Lee
Eugine
E
Department of Theriogenology and Biotechnololgy, College of Veterinary Medicine, Seoul National University, Seoul 151-742, South Korea.
Lee
So Hyun
SH
Kim
Sue
S
Jeong
Yeon Woo
YW
Kim
Ji Hye
JH
Koo
Ok Jae
OJ
Park
Seon Mi
SM
Hashem
M A
MA
Hossein
M S
MS
Son
Hye Young
HY
Lee
Chang Kyu
CK
Hwang
Woo Suk
WS
Kang
Sung Keun
SK
Lee
Byeong Chun
BC
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D019976:Cloning, Organism; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000378:metabolism; D005307:Fertilization in Vitro; D005333:Fetus / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005854:Germ Cells / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine; D013556:Swine, Miniature / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution
D051792:Basic Helix-Loop-Helix Proteins; D050991:GATA4 Transcription Factor; D050814:Octamer Transcription Factor-3; D012333:RNA, Messenger; C097341:helix-loop-helix protein, eHAND