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Efficient imaging of amyloid deposits in Drosophila models of human amyloidoses

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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.

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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).

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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.

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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

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  1. Ina Berg
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  2. K Peter R Nilsson
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  3. Stefan Thor
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  4. Per Hammarström
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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.

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Correspondence to Per Hammarström.

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The authors declare no competing financial interests.

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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

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