Advertisement

Polyamines pp 159-170 | Cite as

Use of (Gyro) Gy and Spermine Synthase Transgenic Mice to Study Functions of Spermine

  • Xiaojing Wang
  • Anthony E. Pegg
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 720)

Abstract

The polyamines putrescine, spermidine, and spermine are essential for mammalian cell growth, ­differentiation, and cell death and have important physiological roles in all tissues. Many of the properties of polyamines that can be demonstrated in vitro are common to all three molecules with differences only in potency. Loss of any of the enzymes needed to make either putrescine or spermidine (which also ­prevent the production of spermine) is lethal, but male mice lacking spermine synthase (SpmS) due to a deletion of part of the X chromosome are viable on the B6C3H background. These mice are termed Gyro (Gy) due to their circling behavior. They have a variety of abnormalities including deafness, neurological problems, small size, and a tendency to early death. They can therefore be used to evaluate the physiological function(s) uniquely provided by spermine. They also provide a potential animal model for Snyder-Robinson syndrome (SRS), a rare human inherited disease due to a loss of SpmS activity. An essential control in experiments using Gy mice is to demonstrate that the abnormal phenotypes exhibited by these mice are abolished by providing replacement spermine and this can be accomplished by breeding with CAG-SMS mice that express SpmS from a ubiquitous promoter. Techniques for identifying, characterizing, and using these mouse strains and limitations of this approach are described in this chapter.

Key words

Spermine Spermine synthase Gy mice CAG-SMS mice 

Notes

Acknowledgements

This work was supported by grant 3R01GM026290-29S1 from the NIH using funds from the NIH Recovery Act.

References

  1. 1.
    Pendeville H, Carpino N, Marine JC, Takahashi Y, Muller M, Martial JA, Cleveland JL (2001) The ornithine decarboxylase gene is essential for cell survival during early murine development. Mol Cell Biol 21:6459–6558CrossRefGoogle Scholar
  2. 2.
    Nishimura K, Nakatsu F, Kashiwagi K, Ohno H, Saito H, Saito T, Igarashi K (2002) Essential role of S-adenosylmethionine decarboxylase in mouse embryonic development. Genes Cells 7:41–47PubMedCrossRefGoogle Scholar
  3. 3.
    Korhonen V-P, Niranen K, Halmekyto M, Pietilä M, Diegelman P, Parkkinen JJ, Eloranta T, Porter CW, Alhonen L, Jänne J (2001) Spermine deficiency resulting from targeted disruption of the spermine synthase gene in embryonic stem cells leads to enhanced sensitivity to antiproliferative drugs. Mol Pharmacol 59:231–238PubMedGoogle Scholar
  4. 4.
    Nilsson J, Gritli-Linde A, Heby O (2000) Skin fibroblasts from spermine synthase-deficient hemizygous gryo male (Gy/Y) mice overproduce spermidine and exhibit increased resistance to oxidative stress but decreased resistance to UV irradiation. Biochem J 352:381–387PubMedCrossRefGoogle Scholar
  5. 5.
    Mackintosh CA, Pegg AE (2000) Effect of spermine synthase deficiency on polyamine biosynthesis and content in mice and embryonic fibroblasts and the sensitivity of fibroblasts to 1, 3-bis(2-chloroethyl)-N-nitrosourea. Biochem J 351:439–447PubMedCrossRefGoogle Scholar
  6. 6.
    Rider JE, Hacker A, Mackintosh CA, Pegg AE, Woster PM, Casero RA Jr (2007) Spermine and spermidine mediate protection against oxidative damage caused by hydrogen peroxide. Amino Acids 33:231–240PubMedCrossRefGoogle Scholar
  7. 7.
    Ikeguchi Y, Mackintosh CA, McCloskey DE, Pegg AE (2003) Effect of spermine synthase on the sensitivity of cells to antitumor agents. Biochem J 373:885–892PubMedCrossRefGoogle Scholar
  8. 8.
    Lyon MF, Scriver CR, Baker LR, Tenenhouse HS, Kronick J, Mandla S (1986) The Gy mutation: another cause of X-linked hypophosphatemia in mouse. Proc Natl Acad Sci USA 83(13):4899–4903PubMedCrossRefGoogle Scholar
  9. 9.
    Grieff M, Whyte MP, Thakker RV, Mazzarella R (1997) Sequence analysis of 139 kb in Xp22.1 containing spermine synthase and 5’ region of PEX. Genomics 44:227–231PubMedCrossRefGoogle Scholar
  10. 10.
    Meyer RA Jr, Henley CM, Meyer MH, Morgan PL, McDonald AG, Mills C, Price DK (1998) Partial deletion of both the spermine synthase gene and the Pex gene in the x-linked hypophosphatemic, Gyro (Gy) mouse. Genomics 48:289–295PubMedCrossRefGoogle Scholar
  11. 11.
    Lorenz B, Francis F, Gempel J, Böddrich AJM, Schmahl W, Schmidt J (1998) Spermine deficiency in Gy mice caused by deletion of the spermine synthase gene. Hum Mol Genet 7:541–547PubMedCrossRefGoogle Scholar
  12. 12.
    Wang X, Ikeguchi Y, McCloskey DE, Nelson P, Pegg AE (2004) Spermine synthesis is required for normal viability, growth and fertility in the mouse. J Biol Chem 49:51370–51375CrossRefGoogle Scholar
  13. 13.
    Wang X, Levic S, Gratton MA, Doyle KJ, Yamoah EN, Pegg AE (2009) Spermine ­synthase deficiency leads to deafness and a profound sensitivity to alpha-difluoromethylornithine. J Biol Chem 284:930–937PubMedCrossRefGoogle Scholar
  14. 14.
    Meyer RA Jr, Meyer MH, Gray RW, Bruns ME (1995) Femoral abnormalities and ­vitamin D metabolism in X-linked hypophosphatemic (Hyp and Gy) mice. J Orthop Res 13:30–40PubMedCrossRefGoogle Scholar
  15. 15.
    Strom T, Francis F, Lorenz B, Böddrich A, Econs M, Lehrach H, Meitinger T (1997) Pex gene deletions in Gy and Hyp mice provide mouse models for X-linked hypophosphatemia. Hum Mol Genet 6:165–171PubMedCrossRefGoogle Scholar
  16. 16.
    Carpinelli MR, Wicks IP, Sims NA, O’Donnell K, Hanzinikolas K, Burt R, Foote SJ, Bahlo M, Alexander WS, Hilton DJ (2002) An ethyl-nitrosourea-induced point mutation in phex causes exon skipping, x-linked hypophosphatemia, and rickets. Am J Pathol 161:1925–1933PubMedCrossRefGoogle Scholar
  17. 17.
    Ikeguchi Y, Wang X, McCloskey DE, Coleman CS, Nelson P, Hu G, Shantz LM, Pegg AE (2004) Characterization of transgenic mice with widespread overexpression of spermine synthase. Biochem J 381:701–707PubMedCrossRefGoogle Scholar
  18. 18.
    Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y (1997) Green mice as a source of ubiquitous green cells. FEBS Lett 407:313–319PubMedCrossRefGoogle Scholar
  19. 19.
    Kato M, Yamanouchi K, Ikawa M, Okabe M, Naito K, Tojo H (1999) Efficient selection of transgenic mouse embryos using EGFP as a marker gene. Mol Reprod Dev 54:43–48PubMedCrossRefGoogle Scholar
  20. 20.
    Pegg AE, Wang X (2009) Mouse models to investigate the function of spermine. Commun Integr Biol 2:271–274PubMedCrossRefGoogle Scholar
  21. 21.
    Cason AL, Ikeguchi Y, Skinner C, Wood TC, Lubs HA, Martinez F, Simensen RJ, Stevenson RE, Pegg AE, Schwartz CE (2003) X-Linked spermine synthase gene (SMS) defect: the first polyamine deficiency syndrome. Eur J Hum Genet 11:937–944PubMedCrossRefGoogle Scholar
  22. 22.
    de Alencastro G, McCloskey DE, Kliemann SE, Maranduba CM, Pegg AE, Wang X, Bertola DR, Schwartz CE, Passos-Bueno MR, Sertie AL (2008) New SMS mutation leads to a striking reduction in spermine ­synthase protein function and a severe form of Snyder-Robinson X-linked recessive ­mental retardation syndrome. J Med Genet 45:539–543PubMedCrossRefGoogle Scholar
  23. 23.
    Becerra-Solano LE, Butler J, Castañeda-Cisneros G, McCloskey DE, Wang X, Pegg AE, Schwartz CE, Sánchez-Corona J, Garcia-Ortiz JE (2009) A missense mutation, p.V132G, in the X-linked spermine synthase gene (SMS) causes Snyder-Robinson syndrome. Am J Med Genet A 149A:328–335PubMedCrossRefGoogle Scholar
  24. 24.
    Schwartz C, Pegg AE (2010) Methods in molecular biology. In: Pegg AE, Casero RA Jr (eds) Polyamine protocols. Humana Press, TotowaGoogle Scholar
  25. 25.
    Seiler N, Knödgen B (1985) Determination of polyamines and related compounds by reversed-phase high-perfomance liquid chromatography: improved separation systems. J Chromatogr 339:45–57CrossRefGoogle Scholar
  26. 26.
    Kabra PM, Lee HK, Lubich WP, Marton LW (1986) Solid-phase extraction and determination of dansyl derivatives of unconjugated and acetylated polyamines by reversed-phase liquid chromatography; improved separation systems for polyamines in cerebrospinal fluid, urine and tissue. J Chromatogr Biomed Appl 380:19–32CrossRefGoogle Scholar
  27. 27.
    Häkkinen MR (2010) Polyamine analysis by LC-MS. In: Pegg AE, Casero RA Jr (eds) Methods in molecular biology. Polyamine protocols. Totowa, Humana PressGoogle Scholar
  28. 28.
    Wiest L, Pegg AE (1998) Assay of spermidine and spermine synthase. In: Morgan DML (ed) Methods in molecular biology. Polyamine protocols, vol 79. Humana Press, Totowa, pp 51–58CrossRefGoogle Scholar
  29. 29.
    Schwartz CE, Stevenson RE, Wang X, Pegg AE (2010) Spermine synthase deficiency resulting in X-linked intellectual disability (Snyder-Robinson syndrome). In: Pegg AE, Casero RA Jr (eds) Methods in molecular biology. Polyamine protocols. Totowa, Humana PressGoogle Scholar
  30. 30.
    Sawicki JA, Morris RJ, Monks B, Sakai K, Miyazaki J-I (1998) A composite CMV-IE enhancer/β-actin promoter is ubiquitously expressed in mouse cutaneous epithelium. Exp Cell Res 244:367–369PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Xiaojing Wang
    • 1
  • Anthony E. Pegg
    • 2
  1. 1.Department of Cellular and Molecular Physiology, Milton S. Hershey Medical CenterPennsylvania State University College of MedicineHersheyUSA
  2. 2.College of Medicine, Milton S. Hershey Medical CenterPennsylvania State UniversityHersheyUSA

Personalised recommendations