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.

  • Protocol
  • Published:

Efficient imaging of amyloid deposits in Drosophila models of human amyloidoses

Nature Protocols volume 5, pages 935–944 (2010)Cite this article

Subjects

Abstract

Drosophila melanogaster is emerging as an important model system for neurodegenerative disease research. In this protocol, we describe an efficient method for imaging amyloid deposits in the Drosophila brain, by the use of a luminescent-conjugated oligothiophene (LCO), p-FTAA polymer probe. We also demonstrate the feasibility of co-staining with antibodies and compare the LCO staining with standard amyloid-specific probes. The LCO protocol enables high-resolution imaging of several different protein aggregates, such as Aβ1-42, Aβ1-42E22G, Transthyretin V30M and human Tau, in the Drosophila brain. Aβ and Tau aggregates could also be distinguished from each other because of distinct LCO emission spectra. Furthermore, this protocol enables three-dimensional brain mapping of amyloid distribution in whole-mount Drosophila brains. The use of p-FTAA combined with other probes, antibodies and/or dyes will aid the rapid characterization of various amyloid deposits in the rapidly growing number of Drosophila models of neurodegenerative diseases.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Staining of 20 μm sections of Drosophila brains.
Figure 2: Sections (20 μm) of Drosophila brains co-stained with p-FTAA and antibody.
Figure 3: Comparison of LCO staining of Drosophila and human tissue.
Figure 4: Composite three-dimensional micrograph of amyloid distribution in whole Drosophila brain of 10-d-old double expressing Aβ1-42 Drosophila line (C155-Gal4/UAS-Aβ1-42; UAS-Aβ1-42).

Similar content being viewed by others

References

  1. Crowther, D.C. et al. Intraneuronal abeta, non-amyloid aggregates and neurodegeneration in a Drosophila model of Alzheimer's disease. Neuroscience 132, 123–135 (2005).

    Article  CAS  PubMed  Google Scholar 

  2. Wittmann, C.W. et al. Tauopathy in Drosophila: neurodegeneration without neurofibrillary tangles. Science 293, 711–714 (2001).

    Article  CAS  PubMed  Google Scholar 

  3. Jackson, G.R. et al. Polyglutamine-expanded human huntingtin transgenes induce degeneration of Drosophila photoreceptor neurons. Neuron 21, 633–642 (1998).

    Article  CAS  Google Scholar 

  4. Pokrzywa, M., Dacklin, I., Hultmark, D. & Lundgren, E. Misfolded transthyretin causes behavioral changes in a Drosophila model for transthyretin-associated amyloidosis. Eur. J. Neurosci. 26, 913–924 (2007).

    Article  PubMed  Google Scholar 

  5. Berg, I., Thor, S. & Hammarstrom, P. Modeling familial amyloidotic polyneuropathy (Transthyretin V30M) in Drosophila melanogaster. Neurodegener. Dis. 6, 127–138 (2009).

    Article  CAS  PubMed  Google Scholar 

  6. Greeve, I. et al. Age-dependent neurodegeneration and Alzheimer-amyloid plaque formation in transgenic Drosophila. J. Neurosci. 24, 3899–3906 (2004).

    Article  CAS  PubMed  Google Scholar 

  7. Iijima, K. et al. Abeta42 mutants with different aggregation profiles induce distinct pathologies in Drosophila. PloS One 3, e1703 (2008).

    Article  PubMed  PubMed Central  Google Scholar 

  8. Nilsson, K.P.R., Herland, A., Hammarstrom, P. & Inganas, O. Conjugated polyelectrolytes: conformation-sensitive optical probes for detection of amyloid fibril formation. Biochemistry 44, 3718–3724 (2005).

    Article  CAS  PubMed  Google Scholar 

  9. Nilsson, K.P.R. et al. Imaging distinct conformational states of amyloid-beta fibrils in Alzheimer's disease using novel luminescent probes. ACS Chem. Biol. 2, 553–560 (2007).

    Article  CAS  PubMed  Google Scholar 

  10. Åslund, A. et al. Novel pentameric thiophene derivatives for in vitro and in vivo optical imaging of a plethora of protein aggregates in cerebral amyloidoses. ACS Chem. Biol. 4, 673–684 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  11. Sigurdson, C.J. et al. Prion strain discrimination using luminescent conjugated polymers. Nat. Meth. 4, 1023–1030 (2007).

    Article  CAS  Google Scholar 

  12. Brand, A.H. & Perrimon, N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118, 401–415 (1993).

    CAS  Google Scholar 

  13. Åslund, A., Nilsson, K.P.R. & Konradsson, P. Synthesis of a pentathiophene fluorescent probe, 4′,3″′-bis-carboxymethyl-[2,2′;5′,2″;5″,2″′;5″′,2″″]quinquethiophene-5,5″″-dicarboxylic acid (p-FTAA). Nat. Protoc. Netw. DOI: 10.1038/nprot.2010.24 (2010).

    Article  CAS  PubMed  Google Scholar 

  14. Lin, D.M. & Goodman, C.S. Ectopic and increased expression of Fasciclin II alters motoneuron growth cone guidance. Neuron 13, 507–523 (1994).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Damian Crowther for the kind gift of the UAS-Aβ1-42E22G and UAS-Aβ1-42 and to Mel B. Feany for the gift of the UAS-Tau flies. We thank the division of Pathology at the University of Linköping for the kind gift of the ToPro3 solution and for the excellent technical support by Magnus Baumgardt and Mattias Alenius at the division of Molecular Genetics at the Univerity of Linköping. A special thank to Andreas Åslund at the division of Organic Chemistry of Linköping University for the contribution of p-FTAA. This work was supported by a generous gift from Astrid and Georg Olsson (P.H. and K.P.R.N.), the Swedish Foundation for Strategic Research (S.T., K.P.R.N. and P.H.), 'Hjärnfonden' (S.T.), The Knut and Alice Wallenberg foundation (S.T., K.P.R.N. and P.H.), and the Swedish Research Council (S.T. and P.H.). P.H. and S.T. are Swedish Royal Academy of Science Research Fellows sponsored by a grant from the Knut and Alice Wallenberg Foundation.

Author information

Authors and Affiliations

  1. IFM-Department of Chemistry, Linköping University, Linköping, Sweden

    Ina Berg, K Peter R Nilsson & Per Hammarström

  2. IKE-Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden

    Stefan Thor

Authors
  1. Ina Berg
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K Peter R Nilsson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stefan Thor
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Per Hammarström
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

I.B., K.P.R.N., S.T. and P.H. designed the research; I.B. carried out the research; I.B., K.P.R.N. and P.H. analyzed data; and I.B. and P.H. wrote the paper.

Corresponding author

Correspondence to Per Hammarström.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Berg, I., Nilsson, K., Thor, S. et al. Efficient imaging of amyloid deposits in Drosophila models of human amyloidoses. Nat Protoc 5, 935–944 (2010). https://doi.org/10.1038/nprot.2010.41

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2010.41

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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