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Molecular & Cellular Proteomics 5:1559-1566, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Proteomic Analysis of the Extraembryonic Tissue from Cloned Porcine Embryos^*,S

Jung-Il Chae^,, Seong-Keun Cho^,¶, Jung-Woo Seo, Tae-Sung Yoon^||, Kyu-Sun Lee, Jin-Hoi Kim^¶, Kyung-Kwang Lee, Yong-Mahn Han^,**, and Kweon Yu^,

From the Centre for Development and Differentiation and ^|| Systemic Proteomics Research Centre, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-333, Korea, ^¶ Division of Applied Life Science, Gyeong-Sang National University, Jinju 660-701, Korea, and ^** Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Korea

Cloned animals developed from somatic cell nuclear transfer (SCNT) embryos are useful resources for agricultural and medical applications. However, the birth rate in the cloned animals is very low, and the cloned animals that have survived show various developmental defects. In this report, we present the morphology and differentially regulated proteins in the extraembryonic tissue from SCNT embryos to understand the molecular nature of the tissue. We examined 26-day-old SCNT porcine embryos at which the sonogram can first detect pregnancy. The extraembryonic tissue from SCNT embryos was abnormally small compared with the control. In the proteomic analysis with the SCNT extraembryonic tissue, 39 proteins were identified as differentially regulated proteins. Among up-regulated proteins, Annexins and Hsp27 were found. They are closely related to the processes of apoptosis. Among down-regulated proteins, Peroxiredoxins and anaerobic glycolytic enzymes were identified. In the Western blot analysis, antioxidant enzymes and the antiapoptotic Bcl-2 protein were down-regulated, and caspases were up-regulated. In the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) assay with the placenta from SCNT embryos, apoptotic trophoblasts were observed. These results demonstrate that a major reason for the low birth rate of cloned animals is due to abnormal apoptosis in the extraembryonic tissue during early pregnancy.

To whom correspondence may be addressed. Tel.: 82-42-860-4642; Fax: 82-42-860-4608; E-mail: kweonyu{at}kribb.re.kr

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