Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Extracellular Superoxide Dismutase (EC-SOD) Transgenic Mice: Possible Animal Model for Various Skin Changes

  • Kim, Sung-Hyun (School of Life Science and Biotechnology, Kyungpook National University) ;
  • Kim, Myoung-Ok (School of Life Science and Biotechnology, Kyungpook National University) ;
  • Lee, Sang-Gyu (School of Life Science and Biotechnology, Kyungpook National University) ;
  • Ryoo, Zae-Young (School of Life Science and Biotechnology, Kyungpook National University)
  • Published : 2006.12.31
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Abstract

We have generated transgenic mice that expressed mouse extracellular superoxide dismutase (EC-SOD) in their skin. In particular, the expression plasmid DNA containing human keratin K14 promoter was used to direct the keratinocyte-specific transcription of the transgene. To compare intron-dependent and intron-independent gene expression, we constructed two vectors. The vector B, which contains the rabbit -globin intron 2, was not effective for mouse EC-SOD overexpression. The EC-SOD transcript was detected in the skin, as determined by Northern blot analysis. Furthermore, EC-SOD protein was detected in the skin tissue, as demonstrated by Western blot analysis. To evaluate the expression levels of EC-SOD in various tissues, we purified EC-SOD from the skin, lungs, brain, kidneys, livers, and spleen of transgenic mice and measured its activities. EC-SOD activities in the transgenic mice skin were approximately 7 fold higher than in wild-type mice. These results suggest that the mouse overexpressing vector not only induces keratinocyte-specific expression of EC-SOD, but also expresses successfully functional EC-SOD. Thus, these transgenic mice appeared to be useful for the expression of the EC-SOD gene and subsequent analysis of various skin changes, such as erythema, inflamation, photoaging, and skin tumors.

Keywords

References

  1. 1. Bowler RP, Arcaroli J, Crapo JD, Ross A, Solt JW, Abraham E (2001): Extracelluar superoxide dismutase attenuates lung injury after hemorrhage. Am J Respir Crit Care Med 164:290-294 https://doi.org/10.1164/ajrccm.164.2.2011054
  2. Buchman AR, Berg P (1988): Comparison of introndependent and intron-independent gene expression. Mol Cell BioI 8:4395-4405 https://doi.org/10.1128/MCB.8.10.4395
  3. Carlsson LM, Jonssen J, Edeund T, Marklund SL (1995): Micelacking extracelluar superoxide dismutase are more sensitive to hyperoxia. Proc Natl Sci VSA 92:6264-6268
  4. Carraro C, Pathak MA (1988): Characterization of superoxide dismutase from mammalian skin epidermis. J Invest Dermatol 90:31-36 https://doi.org/10.1111/1523-1747.ep12462534
  5. Crapo JD, Tierney DF (1974): Superoxide dismutase and pulmonary oxygen toxicity. Am J Physiol 226:1401-1407
  6. Crapo JD, McCord JM Fridovich I (1978): Preparation and assay of superoxide dismutase. Methods Enzymol 53:382-393 https://doi.org/10.1016/S0076-6879(78)53044-9
  7. Fattman CL, Enghild JJ, Crapo JD, Schaefer LM, Valnickova Z, Oury TD (2000): Purification and characterization of extracellular superoxide dismutase in mouse lung. Biochem Biophysic Res Commun 275: 542-548 https://doi.org/10.1006/bbrc.2000.3327
  8. Folz RJ, Guan J, Seldin MF, Oury TD, Enghild JJ, Crapo JD (1997): Mouse extracellular superoxide dismutase: primary structure, tissue-specific gene expression, chromosomal localization, and lung in situ hybridization. Am J Respir Cell Mol BioI 17:393-403 https://doi.org/10.1165/ajrcmb.17.4.2826
  9. Freeman BA, Carpo JD (1982): Biology of disease: Free radicals and tissue injury. Lab Invest 5:412-425
  10. Horinig B, Landmesser U, Kohler C, Ahlersmann D, Spiekerrmann S, Christoph A, Tatge H, Drexler F (2001): Comparatiave effect if ace inhibition and angiotensin II type 1 receptor antagonism on bioavailability of mitric oxide in patients with coronary artery disease: role of superoxide dismutase. Circulation 103:799-805 https://doi.org/10.1161/01.CIR.103.6.799
  11. Jonsson LM, Edlund T, Marklund SL, Sandstrom T (2002): Increased ozone-induced airway neutrophilic inflammation in extracelluar-superoxide dismutase null mice. Respir Med 96:209-214 https://doi.org/10.1053/rmed.2001.1253
  12. Marklund SL (1980): Distribution of Cu/Zn superoxide dismutase and Mn superoxide dismutase in human tissues and extracellular fluids. Acta Physiologica Scandinavia 492:19-23
  13. Marklund SL (1990): Analysis of extracellular superoxide dismutase in tissue homogenates and extracellular fluids. Methods Enzymol 186:260-265 https://doi.org/10.1016/0076-6879(90)86117-E
  14. Marklund SL (2002): Extracellular superoxide dismutase. Methods Enzymol 349:74-80 https://doi.org/10.1016/S0076-6879(02)49322-6
  15. Ookawara T, Imazeki N, Matsubara O, Kizaki T, Ohishi SS, Nako C, Sato Y, Ohno H (1998): Tissue distribution of immunoreactive mouse extracellular superoxide dismutase. Am Physiol Soc C840-C847
  16. Ookawara T, Matsuura N, Ohishi T, Okazaki T, Suzuki K, Hitomi Y, Suzuki K, Ohno H (2000): Serum extracellular superoxide dismutase in pediatric patients with various diseases as judged by an ELISA. Res Commun Mol Pharmacol 107:291-296
  17. Oury TD, Ho YS, Piantadosi CA, Crapo JD (1992): Extracelluar superoxide dismutase, nitric oxide, and central nervous system $O_{2}$ toxicity. Proc Natl Acad Sci USA 89:9715-9719
  18. Oury TD, Crapo JD, Valnickova Z, Enghild J (1996): Human extracellular superoxide dismutase is a tetramer composed of two disulphide-linked dimers: A simplified, high-yield purification of extracellular superoxide dismutase. Biochem J 317:51-57 https://doi.org/10.1042/bj3170051
  19. Pugliese PT (1995): The skin, free radical, and oxidative stress. Dermatology Nursing 7:361-371
  20. Pugliese PT (1998): The skin's antioxidant systems. Dermatology Nursing 10:401-416
  21. Staggers WR, Paterson AJ, Kudlow JE (1995): Sequence of the functional human keratin K14 promoter. Gene 153: 297-298 https://doi.org/10.1016/0378-1119(94)00731-7
  22. Steinman HM, Naik VR, Abernethy JL, Hill RL (1974): Bovine erythrocyte superoxide dismutase: Complete amino acid sequence. J BioI Chem 249:7326-7338
  23. Zelko IN, Mariani TJ, Folz RJ (2002): Superoxide dismutase multigene family: A comparison of the CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression. Free Radic BioI Med 33:337-349 https://doi.org/10.1016/S0891-5849(02)00905-X