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Fluorescent nanodiamond tracking reveals intraneuronal transport abnormalities induced by brain-disease-related genetic risk factors

Nature Nanotechnology volume 12, pages 322–328 (2017)Cite this article

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Abstract

Brain diseases such as autism and Alzheimer's disease (each inflicting >1% of the world population) involve a large network of genes displaying subtle changes in their expression1. Abnormalities in intraneuronal transport have been linked to genetic risk factors found in patients2,3, suggesting the relevance of measuring this key biological process. However, current techniques are not sensitive enough to detect minor abnormalities. Here we report a sensitive method to measure the changes in intraneuronal transport induced by brain-disease-related genetic risk factors using fluorescent nanodiamonds (FNDs). We show that the high brightness, photostability and absence of cytotoxicity4 allow FNDs to be tracked inside the branches of dissociated neurons with a spatial resolution of 12 nm and a temporal resolution of 50 ms. As proof of principle, we applied the FND tracking assay on two transgenic mouse lines that mimic the slight changes in protein concentration (∼30%) found in the brains of patients. In both cases, we show that the FND assay is sufficiently sensitive to detect these changes.

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Figure 1: Intraneuronal transport monitoring by FND tracking.
Figure 2: Transport parameter quantification for anterograde and retrograde motion in axons of neurons grown in a microfluidic chamber.
Figure 3: Effect of nocodazole and Alzheimer's disease-related amyloid-β1–42 oligomers on intraneuronal transport.
Figure 4: Sensitivity of the FND tracking assay to slight concentration changes in the autism-related Mark1 protein.
Figure 5: Sensitivity of the FND tracking assay to the slight overexpression of autism-related Slc25a12.

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Acknowledgements

We thank P. Tchénio for his comments on the manuscript, A. Lombard for his contribution to data processing and J. Lipecka for her help with western blotting. This work was supported by a public grant overseen by the French National research Agency (ANR) as part of the Investissement d'Avenir programme IDEX Paris-Saclay ANR-11-IDEX- 0003-02 (S.H.); the ANR and the Ministry of Science and Technology (Taiwan) through grant ANR-2010-INTB-1002 (M.S., F.T., H.-C.C. and C.-C.W.); the ANR through the Eranet Euronanomed project 2011-ERNA-006 (F.T., M.S., C.S. and A.C.); the Fondation Jérôme Lejeune (M.S.), the European Union FP7-Health (Grant Agreement n° 305299) AgedBrainSysBio grant (M.S.); grant number 12018592 from Region Ile-de-France (F.T.); CNRS Programme Interdisciplinaire (F.T. and M.S.) and Triangle de la Physique contract no. 2012-038T (F.T. and M.S.).

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Author notes

  1. Nitin Mohan, Sophie Massou & Marie-Pierre Adam

    Present address: Present addresses: Institut de Ciències Fotòniques (ICFO), 08860 Castelledefels, Spain (N.M.); Institut Interdisciplinaire de Neurosciences, CNRS, Université de Bordeaux, 33077 Bordeaux, France (S.M.); European Laboratory for Non-Linear Spectroscopy, University of Florence, 50019 Sesto Fiorentino, Italy (M.-P.A.),

  2. François Treussart and Michel Simonneau: These authors contributed equally to this work

Authors and Affiliations

  1. Laboratoire Aimé Cotton, CNRS, Université Paris-Sud, ENS Cachan, Université Paris-Saclay, Orsay, 91405, France

    Simon Haziza, Nitin Mohan, Sophie Massou, Marie-Pierre Adam, Xuan Loc Le, François Treussart & Michel Simonneau

  2. Centre de Psychiatrie et Neurosciences, INSERM U894, Université Paris-Descartes, Paris, 75014, France

    Simon Haziza, Yann Loe-Mie, Aude-Marie Lepagnol-Bestel, Julia Viard, Rachel Daudin, Emilie Dorard, Christiane Rose, Brigitte Potier, Bernadette Allinquant & Michel Simonneau

  3. Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Institut de Génomique, Centre National de Génotypage, Evry, 91057, France

    Christine Plancon

  4. Institut de génétique et de biologie moléculaire et cellulaire, CNRS UMR 7104, INSERM U 964, Université de Strasbourg, Illkirch-Graffenstaden, 67400, France

    Pascale Koebel & Yann Herault

  5. Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, 106, Taiwan

    Feng-Jen Hsieh & Huan-Cheng Chang

  6. Department of Applied Chemistry, National Chi Nan University, Puli, Nantou Hsien, 545, Taiwan

    Chih-Che Wu

  7. Neuroscience Institute, CNR, Milano, 20129, Italy

    Carlo Sala

  8. Department of Psychiatry, Institute of Neuroscience, Trinity College Dublin, Dublin 2, Ireland

    Aiden Corvin

  9. Department of Biology, ENS Cachan, Université Paris-Saclay, Cachan, 94235, France

    Michel Simonneau

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Contributions

S.H., F.T. and M.S. conceived and designed the experiments. S.H., N.M., Y.L.-M., A.-M.L.-B., M.-P.A., S.M., X.L.L., J.V., C.P., E.D., C.R. and R.D. performed the experiments. S.H., N.M., S.M., M.-P.A. and X.L.L. analysed the data. P.K., F.-J.H., C.-C.W., B.P., Y.H., C.S., A.C., B.A. and H.-C.C. contributed to materials and analysis tools. S.H., F.T. and M.S. wrote the paper.

Corresponding authors

Correspondence to Simon Haziza, François Treussart or Michel Simonneau.

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Haziza, S., Mohan, N., Loe-Mie, Y. et al. Fluorescent nanodiamond tracking reveals intraneuronal transport abnormalities induced by brain-disease-related genetic risk factors. Nature Nanotech 12, 322–328 (2017). https://doi.org/10.1038/nnano.2016.260

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