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Bioluminescent Imaging in Bone

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Book cover Osteoporosis

Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 455))

Abstract

Monitoring gene expression in vitro and in vivo, is crucial when analyzing osteogenesis and developing effective bone gene therapy protocols. Until recently, molecular analytical tools were only able to detect protein expression either in vitro or in vivo. These systems include histology and immunohistochemistry, fluorescent imaging, PET (micro-PET), CT (micro-CT), and bioluminescent imaging. The last is the only system to date that can enable efficient quantitative monitoring of gene expression both in vitro and in vivo. Effective bioluminescent imaging in bone can be achieved by using transgenic mice harboring the luciferase reporter gene, downstream of an osteogenesis specific promoter. The aim of this chapter is to comprehensively describe the various protocols needed for the detection of bioluminescence in bone development and repair.

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References

  1. 1. Bar, I., Zilberman, Y., Zeira, E., et al. (2003) Molecular Imaging of the skeleton: quantitative real time bioluminescence in transgenic mice. J Bone Miner Res 18, 570–578.

    Article  CAS  Google Scholar 

  2. 2. Wu, J. C., Sundaresan, G., Iyer, M., et al. (2001) Noninvasive optical imaging of firefly luciferase reporter gene expression in skeletal muscles of living mice. Mol Ther 4, 297–306.

    Article  CAS  PubMed  Google Scholar 

  3. 3. Honigman, A., Zeira, E., Ohana, P., et al. (2001) Imaging transgene expression in live animals. Mol Ther 4, 239–249.

    Article  CAS  PubMed  Google Scholar 

  4. 4. Contag, P. R., Olomu, I. N., Stevenson, D. K., et al. (1998) Bioluminescent indicators in living mammals. Nat Med 4, 245–247.

    Article  CAS  PubMed  Google Scholar 

  5. 5. Lian, J. B., Stein, G. S., Stein, J. L., et al. (1998) Osteocalcin gene promoter: unlocking the secrets for regulation of osteoblast growth an differentiation. J Cell Biochem 30/31, 62–72.

    Article  Google Scholar 

  6. 6. Hauschka, P. V., Lian, J. B., Cole, D. E. C., et al. (1989) Osteocalcin and matrix Gla protein: vitamin K-dependent proteins in bone. Physiol Rev 69, 990–1047.

    CAS  PubMed  Google Scholar 

  7. 7. Sims, N. A., White, C. P., Sunn, K.L., et al. (1997) Human and murine osteocalcin gene expression: conserved tissue restricted expression and divergent responses to 1,25-dihydroxyvitamin D3 in vivo. Mol Endocrinol 11, 1695–1708.

    Article  CAS  PubMed  Google Scholar 

  8. 8. Thomas, G. P., Bourne, A., Eisman, J. A., et al. (2000) Species-divergent regulation of human and mouse osteocalcin genes by calciotropic hormones. Exp Cell Res 258, 395–402.

    Article  CAS  PubMed  Google Scholar 

  9. 9. Zhao, G., Monier-Faugere, M. C., Langub, M. C., et al. (2000) Targeted overexpression of insulin-like growth factor I to osteoblasts of transgenic mice: Increased trabecular bone volume without increased osteoblast proliferation. Endocrinology 141, 2674–2682.

    Article  CAS  PubMed  Google Scholar 

  10. 10. Clemens, T. L., Tang, H., Maeda, S., et al. (1997) Analysis of osteocalcin expression in transgenic mice reveals a species difference in vitamin D regulation of mouse and human osteocalcin genes. J Bone Miner Res 12, 1570–1576.

    Article  CAS  PubMed  Google Scholar 

  11. 11. Lavon, I., Goldberg, I., Amit, S., et al. (2000) High susceptibility to bacterial infection, but no liver dysfunction, in mice compromised for hepatocyte NF-kappaB activation. Nat Med 6, 573–577.

    Article  CAS  PubMed  Google Scholar 

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Zilberman, Y., Gafni, Y., Pelled, G., Gazit, Z., Gazit, D. (2008). Bioluminescent Imaging in Bone. In: Westendorf, J.J. (eds) Osteoporosis. Methods In Molecular Biology™, vol 455. Humana Press. https://doi.org/10.1007/978-1-59745-104-8_18

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  • DOI: https://doi.org/10.1007/978-1-59745-104-8_18

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-828-7

  • Online ISBN: 978-1-59745-104-8

  • eBook Packages: Springer Protocols

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