Lab Anim Res.  2014 Jun;30(2):45-53. 10.5625/lar.2014.30.2.45.

Proteomic analysis of domestic pig pancreas during development using two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry

• Ahn JY
• Kim IY
• Oh SJ
• Hwang HS
• Yi SS
• Kim YN
• Shin JH
• Yoon YS
• Seong JK

• KMID: 1707443
• DOI: http://doi.org/10.5625/lar.2014.30.2.45

Abstract

Pig pancreas may be a therapeutic resource for human diabetic patients. However, this potential is hindered by a lack of knowledge of the molecular events of pig pancreas development. In this study, the embryonic day 60, neonate and 6-month protein profiles of pig pancreas were ascertained at using two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-time of flight mass spectrometry. Twenty four proteins were differentially expressed during pig pancreas development. Among them, 12 spots increased and 7 spots decreased according to development. The expression of 5 protein were highest at birth. Expression of digestive enzymes including trypsin, pancreatic triacylglycerol lipase and pancreatic alpha-amylase was elevated in adults, whereas chymotrypsins were highly expressed in neonates. Proteins that were abundantly expressed during gestation were alpha-1-antitrypsin, alpha-fetoprotein and transferrins. Taken together, we found out that several proteins were significantly up- or down- regulated from pig pancreas based on developmental stage. This study will provide basis for understanding development of pig pancreas.

Keyword

Domestic pig; pancreas; development; 2-DE proteomics; MALDI-TOF

MeSH Terms

Adult
alpha-Amylases
alpha-Fetoproteins
Chymotrypsin
Electrophoresis*
Electrophoresis, Gel, Two-Dimensional
Humans
Infant, Newborn
Lipase
Mass Spectrometry*
Pancreas*
Parturition
Pregnancy
Sus scrofa*
Transferrin
Transferrins
Trypsin
Chymotrypsin
Lipase
Transferrin
Transferrins
Trypsin
alpha-Amylases
alpha-Fetoproteins

Figure

• 

  Figure 1 Representative Coomassie blue stained 2-DE gels of pig pancreas based on developmental stages. The proteins (1 mg) were separated on a 24 cm, pH 3-10 nonlinear strip, followed by an 8-18% gradient SDS-PAGE. Proteins were detected by Coomassie blue staining on the 2-DE images of the master gel. The proteins identified are designated with their accession numbers. (A) E60, (B) neonate, (C) adult pig pancreas. The symbol at (C) indicates development group number (1, 2 and 3) and alphabet order in Table 1. Mr indicates molecular weight of proteins.

• 

  Figure 2 2-DE gel enlargements showing proteins that are highly expressed in the adult stage. Various zymogens (pancreatic alpha-amylase precursor, trypsin precursor and pancreatic triacylglycerol lipase) and protein synthesis related proteins (protein disulfide-isomerase precursor, endoplasmin precursor, 60 kDa heat shock protein and phosphatidylethanolamine-binding protein) are up-regulated. The proteins of interest are indicated by arrows. These proteins were identified by MALDI-TOF MS (A. protein disulfide-isomerase precursor, B. glycine amidinotransferase, C. endoplasmin precursor, D. pancreatic alpha-amylase precursor, E. MHC class II antigen, F. trypsin precursor, G. pancreatic triacylglycerol lipase, H. phosphatidylethanolamine-binding protein, I. translationally-controlled tumor protein, J. 60 kDa heat shock protein, K. hypothetical protein FLJ39175, and L. laminin receptor).

• 

  Figure 3 2-DE gel enlargements showing the proteins especially expressed in the neonatal stage. Chymotrypsinogen 2 and 14-3-3 protein are significantly up-regulated at birth. The proteins of interest are indicated by arrows. These proteins were identified by MALDI-TOF MS (A. keratin, 53K type II cytoskeletal, B. chymotrypsinogen 2, C. 14-3-3 protein, D. pyruvate kinase (EC 2.7.1.40) isozyme M2, E. spermidine synthase (EC 2.5.1.16)).

• 

  Figure 4 2-DE gel enlargements showing the proteins especially more expressed in the embryonic stage. Apolipoprotein A-I precursor, alpha-fetoprotein, transferrin and alpha-1-antitrypsin precursor are markedly expressed in fetal developing pancreas. The proteins of interest are indicated by arrows. These proteins were identified by MALDI-TOF MS (A. stathmin (Phosphoprotein p19), B. apolipoprotein A-I precursor, C. alpha-fetoprotein, D. transferrin. E. F25B3.3 kinase like protein (Fragment), F. peptidyl-prolyl cis-trans isomerase A, G. alpha-1-antitrypsin precursor).

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