Lab Anim Res.  2014 Mar;30(1):1-7. 10.5625/lar.2014.30.1.1.

Proteomic analysis of pancreas 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.
  • 2Laboratory of Developmental Biology and Genetics, College of Veterinary Medicine, BK21 Plus Program for Creative Science Research, BIO-MAX institute, Seoul National University, Seoul, Korea. snumouse@snu.ac.kr
  • 3Department of Pathology and Laboratory of Immune Regulation, College of Medicine, Seoul National University, Seoul, Korea.
  • 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.

Abstract

Organ transplantation is limited by the shortage of human organs. Many studies have sought to overcome this hurdle by using animal organs. Porcine organs, especially from miniature pigs, have been used for organ xenotransplantation rather than nonhuman primates. While the molecular profiling for transplantation is well known in humans and rodents, the situation for pigs is almost completely unknown. The present study examined protein regulation of the developing stages of the pancreatic proteome (4 day-old miniature neonate, 19 day-old miniature piglet, and 14 month-old miniature adult pigs) using two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-time of flight mass spectrometry. Thirteen different expressed spots were observed and nine were identified. The data presented within this study provides critical direction relating to the development of pancreas of miniature pigs, which will assist future proteome analysis of the pancreas, and advance our understanding of the hurdles facing xenotransplantation.

Keyword

Human organ; pancreas; miniature pig; proteome; 2-DE MALDI-TOF

MeSH Terms

Adult
Animal Structures
Electrophoresis*
Electrophoresis, Gel, Two-Dimensional
Humans
Infant
Infant, Newborn
Mass Spectrometry*
Organ Transplantation
Pancreas*
Primates
Proteome
Rodentia
Swine*
Transplantation, Heterologous
Transplants
Proteome

Figure

  • Figure 1 Representative 2-DE gels of pancreas of 4-day-old, 19-day-old and 14-month-old miniature pigs by developmental stages, visualized by Commassie blue staining. Samples of 1mg protein were separated on pH3-10 non-linear IPG strip (24cm) following by 8-18% gradient SDS-PAGE in 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 in the 4-day-old miniature neonate pig, 19-day-old miniature piglet and 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 dot circles show down-expressed patterns. The expression patterns of the different three stages are presented as simple line graphs. Relative optical (ROD) are indicated as relative numbers under the lines compared with the lowest protein optical density (the lowest optical density is made as an '1') of each spot.


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