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Comprehensive mapping of neurotransmitter networks by MALDI–MS imaging

Nature Methods volume 16, pages 1021–1028 (2019)Cite this article

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Abstract

We present a mass spectrometry imaging (MSI) approach for the comprehensive mapping of neurotransmitter networks in specific brain regions. Our fluoromethylpyridinium-based reactive matrices facilitate the covalent charge-tagging of molecules containing phenolic hydroxyl and/or primary or secondary amine groups, including dopaminergic and serotonergic neurotransmitters and their associated metabolites. These matrices improved the matrix-assisted laser desorption/ionization (MALDI)–MSI detection limit toward low-abundance neurotransmitters and facilitated the simultaneous imaging of neurotransmitters in fine structures of the brain at a lateral resolution of 10 µm. We demonstrate strategies for the identification of unknown molecular species using the innate chemoselectivity of the reactive matrices and the unique isotopic pattern of a brominated reactive matrix. We illustrate the capabilities of the developed method on Parkinsonian brain samples from human post-mortem tissue and animal models. The direct imaging of neurotransmitter systems provides a method for exploring how various neurological diseases affect specific brain regions through neurotransmitter modulation.

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Fig. 1: Polyphenylated fluoromethylpyridinium reactive matrix enables improved detection and high lateral resolution imaging of neurotransmitters and their metabolites.
Fig. 2: Identification of unknown compounds using different alkylpyridinium reactive matrices (FMP-10 and FMP-9), a brominated reactive matrix (FMP-8), and/or multiple orders of derivatization.
Fig. 3: MALDI-MS images of neurotransmitters and metabolites acquired from a tissue section of a unilateral 6-OHDA lesioned Parkinson’s disease model rat treated with L-DOPA.
Fig. 4: MALDI-MS images and relative quantitation of neurotransmitters and metabolites acquired from a tissue section of an L-DOPA-d3 treated unilateral 6-OHDA lesioned Parkinson’s disease model rat.
Fig. 5: MALDI–MS images of neurotransmitters and other biologically active substances in coronal primate Parkinson’s disease model brain tissue sections.
Fig. 6: MALDI-MS images of neurotransmitters and associated metabolites in the striatum of a parkinsonian human brain tissue section.

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Data availability

The MALDI–MSI datasets that support the findings of this study are available from the corresponding author upon request. NMR data for all compounds is available in the Supplementary Note. MS and MS/MS spectra for derivatized standards are available in the supplementary information. Source data are available online for Fig. 4.

Code availability

Quantitative data analyses were performed using the msIQuant imaging software (v.2.0.1.15). The software is freely available at https://ms-imaging.org/wp/paquan/.

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Acknowledgements

This work was supported by the Swedish Brain Foundation (grant FO2018-0292 to P.E.A.), the Swedish Foundation for Strategic Research (grant RIF14-0078 to P.E.A), the European FP7 ITN Community’s Seventh Framework Program ARIADME (grant 607517 to P.E.A.), the Swedish Research Council, Medicine and Health (grant 2018–03320 to P.E.A.), the Swedish Research Council, Natural and Engineering Science (grant 2018–05501 to P.E.A. and 2018–05133 to L.R.O.) and the Science for Life Laboratory (to P.E.A.). Confocal microscope images were acquired at the BioVis core facility at Uppsala University with help of the BioVis staff. We thank M. Witt for support using the 12T FTICR MS instrument and S. Dengler for experimental assistance.

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Authors and Affiliations

  1. Medical Mass Spectrometry Imaging, Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden

    Mohammadreza Shariatgorji, Anna Nilsson, Elva Fridjonsdottir, Theodosia Vallianatou, Patrik Källback & Per E. Andrén

  2. Science for Life Laboratory, National Resource for Mass Spectrometry Imaging, Uppsala University, Uppsala, Sweden

    Mohammadreza Shariatgorji, Anna Nilsson & Per E. Andrén

  3. Department of Medicinal Chemistry, Uppsala University, Uppsala, Sweden

    Luay Katan, Jonas Sävmarker & Luke R. Odell

  4. Section of Neurology, Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden

    Ioannis Mantas, Xiaoqun Zhang & Per Svenningsson

  5. Institut des Maladies Neurodégénératives, Université de Bordeaux, Bordeaux, France

    Erwan Bezard

  6. Institut des Maladies Neurodégénératives, CNRS, Bordeaux, France

    Erwan Bezard

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Contributions

M.S. and A.N. conceived the methodology, designed experiments, acquired, analyzed and interpreted data, and wrote the manuscript; E.F. acquired and analyzed data and wrote part of the manuscript; T.V. acquired and analyzed data; P.K. analyzed data; L.K. and J.S. synthesized the reactive matrices; I.M. and X.Z. performed animal experiments and provided tissue samples; E.B. and P.S. provided tissue samples, supervised animal experiments and edited the manuscript; L.R.O. conceived the methodology, designed experiments, synthesized the reactive matrices, interpreted data and edited the manuscript; P.E.A. conceived the methodology, designed experiments, interpreted data, wrote the manuscript and is principal investigator for the grants that fund this research.

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Correspondence to Luke R. Odell or Per E. Andrén.

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The authors declare no competing interests.

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Peer review information Nina Vogt and Allison Doerr were the primary editors on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team

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Supplementary Information

Supplementary Figs. 1–14 and Supplementary Tables 1–2

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Supplementary Note

Supplementary Video

MALDI-MS imaging of the nigrostriatal dopaminergic pathway. The nigrostriatal pathway is visualized changing the color scaling for ion intensity of the FMP-10 derivatized DA signal (m/z 674.28). MALDI-MSI experiment was replicated four times independently from different animals with similar results.

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Shariatgorji, M., Nilsson, A., Fridjonsdottir, E. et al. Comprehensive mapping of neurotransmitter networks by MALDI–MS imaging. Nat Methods 16, 1021–1028 (2019). https://doi.org/10.1038/s41592-019-0551-3

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