Lab Anim Res.  2013 Sep;29(3):162-167.

Proteomic analysis of liver in miniature pigs according to developmental stages using two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry

Affiliations
  • 1Department of Biomedical Laboratory Science, College of Biomedical Sciences, Soonchunhyang University, Asan, Korea.
  • 2Department of Pathology and Laboratory of Immune Regulation, College of Medicine, Seoul National University, Seoul, Korea.
  • 3Laboratory of Developmental Biology and Genetics, College of Veterinary Medicine, Seoul National University, Seoul, Korea. snumouse@snu.ac.kr
  • 4Transitional Research Institute, College of Medicine, Seoul National University, Seoul, Korea.
  • 5Biomedical Center for Animal Resource and Development, Bio-Max Institute, Seoul National University, Seoul, Korea.
  • 6Department of Biochemistry, Hanyang University, Ansan, Korea.
  • 7Interdisciplinary Program for Bioinformatics, Program for Cancer Biology and Bio-Max Institute, Seoul National University, Seoul, Korea.

Abstract

Due to the shortage of human organ donors for transplant, various studies of xenotransplantation, or the use of animal organs instead of human organs, have been carried out. The organs of porcine are thought to be safer and of a more suitable size for xenotransplantationthan those of nonhuman primates. Understanding the levels of expression of proteins, and their post-translational regulation, would be very practical between different species and among developing stages, though the molecular profiling for xenotransplantation has been rarely studied for porcine, while that of human and rodent is well known. Here, in this present study, we report protein regulation of the developing stages of liver (4-day old neonate, 19-day old piglet and 14-month old adult miniature pigs) using 2-DE and MALDI-TOF. From images of the three different stages, a total of 8 spotswhich were differently regulated were identified, and 5 spots were identified with MALDI-TOF MS. The data presented within this study provides critical direction relating to the development of livers of miniature pigs, which will assist future proteome analysis of the liver, and advance our understanding of the hurdles facing xenotransplantaion.

Keyword

Liver; MALDI-TOF; miniature pig; proteome; 2-DE; xenotransplantation

MeSH Terms

Adult
Animal Structures
Electrophoresis
Humans
Infant, Newborn
Liver
Mass Spectrometry
Primates
Proteins
Proteome
Rodentia
Swine
Tissue Donors
Transplantation, Heterologous
Transplants
Proteins
Proteome

Figure

  • Figure 1 Representative 2-DE gels of livers 4-day, 19-day and 14-month old miniature pigs by developmental stages, which is visualized by Commassie blue staining. Samples of 1 mg protein were separated on pH3-10 non-linear IPG strip (24 cm) following by 8-18% gradient SDS-PAGE gels as the second dimension. Proteins were detected by Coomassie brilliant blue G-250 and compared using ImageMaster™ 2D Platinum Software version 5.0; A: 2-DE gel of 4-day old miniature pig (neonate), B: 2-DE gel of 19-day old miniature pig, C: 2-DE gel of 14-month old miniature pig. The closed circles indicate 13 differentially expressed protein spots among 4-day old miniature neonate pig, 19-day old miniature piglet, 14-month old miniature adult pig.

  • Figure 2 Two-DE images of alterative spots of pancreas according to developmental stage. The circles indicate differentially expressed protein spots and the expression patterns of the different three stages were described as simple line graphs.


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