45 ANALYSIS OF DIFFERENTIALLY EXPRESSED PROTEINS IN THE PLACENTA OF SOMATIC CELL CLONED AND ARTIFICIAL INSEMINATION PIG PLACENTA USING PROTEOMICS
S.Y. Lee A , S.K. Cho A , M.R. Park A , E.K. Lee A and J.H. Kim AADivision of Applied Life Science, College of Agriculture, Gyeongsang National University, Chinju, 660-701, South Korea. Email: salanghae2@hanmail.net
Reproduction, Fertility and Development 17(2) 172-172 https://doi.org/10.1071/RDv17n2Ab45
Submitted: 1 August 2004 Accepted: 1 October 2004 Published: 1 January 2005
Abstract
Somatic cell cloning has been hampered by biological and technical problems. A major limitation of cloning procedures is the extreme inefficiency of producing healthy offspring for reasons such as sudden intrauterine unexplained death and infant death syndrome. In this study, we analyzed differentially expressed protein profiles in the placenta of somatic cell cloned (SCNT) and artificial insemination (AI; control) pigs by proteomics. Protein expression pattern of placentas was established in the pH range 4–7, IEF cell system, and 7.5–17.5% gradient 2-dimensional polyacrylamide gel electrophoresis. Image analysis following silver staining used the spot and PDQuest analysis system. Peptide mass fingerprinting with matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and SWISS-PROT database search were utilized to identify proteins. Around 1000 protein spots were recognized by image analysis on each placenta. In SCNT pig placentas, 37 spots were changed compared with those of AI pig placentas. Among them 11 proteins such as serum albumin precursor and annexin A5 showed an increased protein expression level whereas the expression level of 26 other proteins such as aldose reductase and tropomyosin decreased. Annexin A5 and aldose reductase are implicated in the apoptosis process: the former is an apoptotic marker of oxidative stress; on the other hand, the latter is a critical regulator of TNF-α-induced apoptotic signaling in endothelial cells. Also, tropomyosin is implicated in stabilizing cytoskeleton actin filaments. The expression of these proteins was confirmed by western blot and real-time PCR analysis. These results suggest expression of abnormal placental protein and apoptotic-related protein in SCNT pig placentas affects the regulation of placental growth and development.
This work was supported in part by a grant program from RDA(Biogreen21) and Cho-A, Republic of Korea.
