title: Aberrant gene expression patterns in extraembryonic tissue from cloned porcine embryos.
abstract: The abnormal development of embryos reconstructed by somatic cell nuclear transfer (SCNT) is considered to be associated with consequent changes in gene expression following errors in epigenetic reprogramming. In this study, we carried out SCNT using donor fibroblast cells derived from 3-way hybrids (Landrace×Duroc×Yorkshire). A total of 655 SCNT embryos were transferred, and 6.97±2.3 cloned fetuses were successfully recovered from three surrogates at gestational day 30. An analysis of the 6.97±2.3 cloned embryos revealed that most had severe extraembryonic defects. The extraembryonic tissue from the SCNT embryos was abnormally small compared with that of the control. To investigate the differentially expressed genes between the SCNT and control extraembryonic tissues, we compared the gene expression profiles of the extraembryonic tissues from gestational day 30 cloned pig embryos with those from the control using an annealing control primer-based GeneFishing polymerase chain reaction. As a result, we found that a total of 50 genes were differentially expressed by utilizing 120 ACPs, 38 genes of which were known. Among them, 26 genes were up-regulated, whereas 12 genes were down-regulated. Real-time RT-PCR showed that apoptosis-related genes were expressed significantly higher in SCNT extraembryonic tissue than in the control, whereas metabolism-related genes were expressed at significantly lower levels in the SCNT extraembryonic tissue. These observations strongly indicate that early gestational death of SCNT embryo is caused, at least in part, by the disruption of developing extraembryonic tissues as a result of aberrant gene expression, which results in abnormal apoptosis and metabolism.
authors.affiliation: Division of Animal Biotechnology, National Institute of Animal Science, RDA, Suwon 441-706, Republic of Korea.
authors.forename: Mi-Ryung
authors.initials: MR
authors.lastname: Park
authors.forename: Gi-Sun
authors.initials: GS
authors.lastname: Im
authors.forename: Sung Woo
authors.initials: SW
authors.lastname: Kim
authors.forename: Seongsoo
authors.initials: S
authors.lastname: Hwang
authors.forename: Jae-Hong
authors.initials: JH
authors.lastname: Park
authors.forename: Hyun
authors.initials: H
authors.lastname: Kim
authors.forename: Yoon Jung
authors.initials: YJ
authors.lastname: Do
authors.forename: Soo Bon
authors.initials: SB
authors.lastname: Park
authors.forename: Bo-Suck
authors.initials: BS
authors.lastname: Yang
authors.forename: Young Min
authors.initials: YM
authors.lastname: Song
authors.forename: Jae-Hyeon
authors.initials: JH
authors.lastname: Cho
authors.forename: Yeoung-Gyu
authors.initials: YG
authors.lastname: Ko
mesh_terms: D000818:Animals; D019976:Cloning, Organism / Q000662:veterinary; D005321:Extraembryonic Membranes / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression / Q000502:physiology*; D020869:Gene Expression Profiling / Q000662:veterinary; D006825:Hybridomas / Q000378:metabolism; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D013552:Swine / Q000196:embryology; D059467:Transcriptome