Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

In vivo detection of amyloid-β deposits by near-infrared imaging using an oxazine-derivative probe

Nature Biotechnology volume 23, pages 577–583 (2005)Cite this article

Abstract

As Alzheimer's disease pathogenesis is associated with the formation of insoluble aggregates of amyloid β-peptide, approaches allowing the direct, noninvasive visualization of plaque growth in vivo would be beneficial for biomedical research. Here we describe the synthesis and characterization of the near-infrared fluorescence oxazine dye AOI987, which readily penetrates the intact blood-brain barrier and binds to amyloid plaques. Using near-infrared fluorescence imaging, we demonstrated specific interaction of AOI987 with amyloid plaques in APP23 transgenic mice in vivo, as confirmed by postmortem analysis of brain slices. Quantitative analysis revealed increasing fluorescence signal intensity with increasing plaque load of the animals, and significant binding of AOI987 was observed for APP23 transgenic mice aged 9 months and older. Thus, AOI987 is an attractive probe to noninvasively monitor disease progression in animal models of Alzheimer disease and to evaluate effects of potential Alzheimer disease drugs on the plaque load.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: General synthesis scheme.
Figure 2: Fluorescence characterization and in vitro binding studies.
Figure 3: Concentration of AOI987 (dye 3a) in brain and plasma of anesthetized wild-type mice (C57BL/6Jico) at various time points (5–120 min) after a single intravenous administration of 3 mg/kg.
Figure 4: In vivo imaging of amyloid-β deposits.
Figure 5
Figure 6: Semi-quantitative in vivo imaging of amyloid-β deposits in living mice.

References

  1. Meyer-Luehmann, M. et al. Extracellular amyloid formation and associated pathology in neuronal grafts. Nat. Neurosci. 6, 1–8 (2003).

    Article  Google Scholar 

  2. Nussbaum, R.L. & Ellis, C.E. Alzheimer's disease and Parkinson's disease. N. Engl. J. Med. 348, 1356–1364 (2003).

    Article  CAS  Google Scholar 

  3. Hardy, J. & Selkoe, D.J. The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeutics. Science 297, 353–356 (2002).

    Article  CAS  Google Scholar 

  4. Zamrini, E., De Santi, S. & Tolar, M. Imaging is superior to cognitive testing for early diagnosis of Alzheimer's disease. Neurobiol. Aging 25, 685–691 (2004).

    Article  Google Scholar 

  5. Benveniste, H., Einstein, G., Kim, K.R., Hulette, C. & Johnson, G.A. Detection of neuritic plaques in Alzheimer's disease by magnetic resonance microscopy. Proc. Natl. Acad. Sci. USA 96, 14079–14084 (1999).

    Article  CAS  Google Scholar 

  6. Zhang, J. et al. Detection of amyloid plaques in mouse models of Alzheimer's disease by magnetic resonance imaging. Magn. Reson. Med. 51, 452–457 (2004).

    Article  Google Scholar 

  7. Mathis, C.A., Wang, Y. & Klunk, W.E. Imaging β-amyloid plaques and neurofibrillary tangles in the aging human brain. Curr. Pharm. Des. 10, 1469–1492 (2004).

    Article  CAS  Google Scholar 

  8. Bacskai, B.J., Klunk, W.E., Mathis, C.A. & Hyman, B.T. Imaging amyloid-β deposits in vivo. J. Cereb. Blood Flow Metab. 22, 1035–1041 (2002).

    Article  CAS  Google Scholar 

  9. Ono, M. et al. 11C-labeled stilbene derivatives as Aβ-aggregate-specific PET imaging agents for Alzheimer's disease. Nucl. Med. Biol. 30, 565–571 (2003).

    Article  CAS  Google Scholar 

  10. Suemoto, T. et al. In vivo labeling of amyloid with BF-108. Neurosci. Res. 48, 65–74 (2004).

    Article  CAS  Google Scholar 

  11. Okamura, N. et al. Styrylbenzoxazole derivatives for in vivo imaging of amyloid plaques in the brain. J. Neurosci. 24, 2535–2541 (2004).

    Article  CAS  Google Scholar 

  12. Klunk, W.E. et al. Uncharged thioflavin-T derivatives bind to amyloid-beta protein with high affinity and readily enter the brain. Life Sci. 69, 1471–1484 (2001).

    Article  CAS  Google Scholar 

  13. Mathis, C.A. et al. Synthesis and evaluation of 11C-labeled 6-substituted 2-arylbenzothiazoles as amyloid imaging agents. J. Med. Chem. 46, 2740–2754 (2003).

    Article  CAS  Google Scholar 

  14. Mathis, C.A. et al. A lipophilic thioflavin-T derivative for positron emission tomography (PET) imaging of amyloid in brain. Bioorg. Med. Chem. Lett. 12, 295–298 (2002).

    Article  CAS  Google Scholar 

  15. Agdeppa, E.D. et al. Binding characteristics of radiofluorinated 6-dialkylamino-2-naphthylethylidene derivatives as positron emission tomography imaging probes for β-amyloid plaques in Alzheimer's disease. J. Neurosci. 21, RC189 1–5 (2001).

    Article  Google Scholar 

  16. Agdeppa, E.D. et al. 2-Dialkylamino-6-acylmalononitrile substituted naphthalenes (DDNP analogs): novel diagnostic and therapeutic tools in Alzheimer's disease. Mol. Imaging Biol. 5, 404–417 (2003).

    Article  Google Scholar 

  17. Wadghiri, Y.Z. et al. Detection of Alzheimer's amyloid in transgenic mice using magnetic resonance microimaging. Magn. Reson. Med. 50, 293–302 (2003).

    Article  CAS  Google Scholar 

  18. Poduslo, J.F. et al. Design and chemical synthesis of a magnetic resonance contrast agent with enhanced in vitro binding, high blood-brain barrier permeability, and in vivo targeting to Alzheimer's disease amyloid plaques. Biochemistry 43, 6064–6075 (2004).

    Article  CAS  Google Scholar 

  19. Poduslo, J.F. et al. Molecular targeting of Alzheimer's amyloid plaques for contrast-enhanced magnetic resonance imaging. Neurobiol. Dis. 11, 315–329 (2002).

    Article  CAS  Google Scholar 

  20. Bacskai, B.J. et al. Imaging of amyloid-β deposits in brains of living mice permits direct observation of clearance of plaques with immunotherapy. Nat. Med. 7, 369–372 (2001).

    Article  CAS  Google Scholar 

  21. Klunk, W. et al. Imaging Aβ plaques in living transgenic mice with multiphoton microscopy and methoxy-X04, a systemically administered Congo red derivative. J. Neuropathol. Exp. Neurol. 61, 797–805 (2002).

    Article  CAS  Google Scholar 

  22. Bacskai, B.J. et al. Four-dimensional multiphoton imaging of brain entry, amyloid binding, and clearance of an amyloid-β ligand in transgenic mice. Proc. Natl. Acad. Sci. USA 100, 12462–12467 (2003).

    Article  CAS  Google Scholar 

  23. Styren, S.D., Hamilton, R.L., Styren, G.C. & Klunk, W.E. X-34, a fluorescent derivative of Congo red: a novel histochemical stain for Alzheimer's disease pathology. J. Histochem. Cytochem. 48, 1223–1232 (2000).

    Article  CAS  Google Scholar 

  24. Shoghi-Jadid, K. et al. Localization of neurofibrillary tangles and beta-amyloid plaques in the brains of living patients with Alzheimer disease. Am. J. Geriatr. Psychiatry 10, 24–35 (2002).

    Article  Google Scholar 

  25. Klunk, W.E. et al. Imaging brain amyloid in Alzheimer's disease with Pittsburgh compound-B. Ann. Neurol. 55, 306–319 (2004).

    Article  CAS  Google Scholar 

  26. Weissleder, R. & Ntziachristos, V. Shedding light onto live molecular targets. Nat. Med. 9, 123–128 (2003).

    Article  CAS  Google Scholar 

  27. Ntziachristos, V., Bremer, C. & Weissleder, R. Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging. Eur. Radiol. 13, 195–208 (2003).

    PubMed  Google Scholar 

  28. Funovics, M., Weissleder, R. & Tung, C.-H. Protease sensors for bioimaging. Anal. Bioanal. Chem. 377, 956–963 (2003).

    Article  CAS  Google Scholar 

  29. Sturchler-Pierrat, C. et al. Two amyloid precursor protein transgenic mouse models with Alzheimer disease-like pathology. Proc. Natl. Acad. Sci. USA 94, 13287–13292 (1997).

    Article  CAS  Google Scholar 

  30. Neuweiler, H. et al. Detection of individual p53-autoantibodies by using quenched peptide-based molecular probes. Angew. Chem. Int. Edn. Engl. 41, 4769–4773 (2002).

    Article  CAS  Google Scholar 

  31. Bremer, C., Tung, C.H. & Weissleder, R. In vivo molecular target assessment of matrix metalloproteinase inhibition. Nat. Med. 7, 743–748 (2001).

    Article  CAS  Google Scholar 

  32. Wunder, A. et al. In vivo imaging of protease activity in arthritis: a novel approach for monitoring treatment response. Arthritis Rheum. 50, 2459–2465 (2004).

    Article  CAS  Google Scholar 

  33. Graves, E.E. et al. A submillimeter resolution fluorescence molecular imaging system for small animal imaging. Med. Phys. 30, 901–911 (2003).

    Article  CAS  Google Scholar 

  34. Rudin, M. & Weissleder, R. Molecular imaging in drug discovery and development. Nat. Rev. Drug Discov. 2, 123–131 (2003).

    Article  CAS  Google Scholar 

  35. Daves, G. & McMurtrey, K.D. Koenig's adducts of N-alkyl(aryl)-aminoethanols and quinones. 3,4-Dihydro-4-alkyl(aryl)-8a-hydroxy-2H–1,4-benzoxazine-6(8aH)-ones. J. Org. Chem. 35, 4252–4253 (1970).

    CAS  Google Scholar 

  36. Klunk, W.E. & Pettegrew, J.W. Development of small molecule probes for the beta-amyloid protein of Alzheimer's disease. Neurobiol. Aging 15, 691–698 (1994).

    Article  CAS  Google Scholar 

  37. Levine, H. III Thioflavin T interaction with synthetic Alzheimer's disease β-amyloid peptides: detection of amyloid aggregation in solution. Protein Sci. 2, 404–410 (1993).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Alexandra Suter for her excellent technical assistance.

Author information

Authors and Affiliations

  1. Discovery Technologies, Novartis Institutes for Biomedical Research, Basel, CH-4002, Switzerland

    Martin Hintersteiner, Willy Kinzy, Rainer Kneuer, Markus Rudin, Markus Stoeckli & Hans-Ulrich Gremlich

  2. Nervous System Department, Novartis Institutes for Biomedical Research, Basel, CH-4002, Switzerland

    Albert Enz, Peter Frey, Anne-Lise Jaton, Ulf Neumann, Matthias Staufenbiel & Karl-Heinz Wiederhold

Authors
  1. Martin Hintersteiner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Albert Enz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter Frey
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anne-Lise Jaton
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Willy Kinzy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rainer Kneuer
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ulf Neumann
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Markus Rudin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Matthias Staufenbiel
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Markus Stoeckli
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Karl-Heinz Wiederhold
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Hans-Ulrich Gremlich
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hans-Ulrich Gremlich.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Hintersteiner, M., Enz, A., Frey, P. et al. In vivo detection of amyloid-β deposits by near-infrared imaging using an oxazine-derivative probe. Nat Biotechnol 23, 577–583 (2005). https://doi.org/10.1038/nbt1085

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1085

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing