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Expression and Secretion of the Insulin-like Growth Factor System Components by Pig Liver Cells

  • Kim, I. (Regional Animal Industry Center, Jinju National University) ;
  • Jin, E.J. (Regional Animal Industry Center, Jinju National University) ;
  • Baik, K. (Regional Animal Industry Center, Jinju National University) ;
  • Park, C.H. (Department of Anatomy and Neurobiology, Institute of Health Science, College of Medicine, Gyeongsang National University) ;
  • Kim, W.K. (Division of Life Science and Genetic Engineering, College of Life and Environmental Sciences, Korea University) ;
  • Kang, C.W. (Department of Physiology, College of Veterinary Medicine, Chonbuk National University) ;
  • Ko, Y. (Division of Life Science and Genetic Engineering, College of Life and Environmental Sciences, Korea University) ;
  • Jang, I. (Regional Animal Industry Center, Jinju National University) ;
  • Choi, W.S. (Department of Anatomy and Neurobiology, Institute of Health Science, College of Medicine, Gyeongsang National University) ;
  • Lee, C.Y. (Regional Animal Industry Center, Jinju National University)
  • Received : 2007.09.20
  • Accepted : 2008.03.07
  • Published : 2008.09.01

Abstract

The aim of the present study was to delineate the expression and secretion of insulin-like growth factor (IGF) system components by pig liver cells. Hepatocytes were prepared from 3-wk-old weanling piglets following a two-step collagenase perfusion procedure, after which the cells were incubated for 24 or 48 h at a density of $2{\pm}10^5$ cells per 35-mm dish in 2-ml Williams' medium E. The cells were found to express the genes encoding IGF-I, IGF-binding proteins (IGFBPs)-2 and -3 and acid-labile subunit (ALS) by reverse transcription-polymerase chain reaction (RT-PCR) following the culture. However, IGF-I was localized to hepatocytes by immunohistochemical analysis, whereas IGFBP-3 was localized to endothelial cells, but not to hepatocytes. This indicated that the IGFBP-3 gene expression detected by RT-PCR was likely to have been contributed by unidentified non-parenchymal cells that had not been removed during the hepatocyte preparation. The conditioned culture medium (CCM) of the cells contained immunoreactive IGF-I and IGF-II, with the latter being seven-fold more abundant than the former. The CCM also contained 43-, 40-, 34-, 31-kDa doublet and 26-kDa IGFBPs as examined by Western ligand blotting. The 40-, 34- and 31-kDa doublet IGFBPs were approximately three-fold as abundant as the 43- and 26-kDa IGFBPs. Moreover, the 43- and 40-kDa doublet and the 34-kDa IGFBPs were immunoprecipitable with IGFBP-3 and IGFBP-2 antibodies, respectively. Overall, these results are similar to those known in the rat, which suggests that the IGF system components are likely to be expressed and secreted in pig liver in a manner similar to that in rat liver.

Keywords

References

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