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Co-aggregation of RNA binding proteins in ALS spinal motor neurons: evidence of a common pathogenic mechanism

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Abstract

While the pathogenesis of amyotrophic lateral sclerosis (ALS) remains to be clearly delineated, there is mounting evidence that altered RNA metabolism is a commonality amongst several of the known genetic variants of the disease. In this study, we evaluated the expression of 10 ALS-associated proteins in spinal motor neurons (MNs) in ALS patients with mutations in C9orf72 (C9orf72GGGGCC-ALS; n = 5), SOD1 (mtSOD1-ALS; n = 9), FUS/TLS (mtFUS/TLS-ALS; n = 2), or TARDBP (mtTDP-43-ALS; n = 2) and contrasted these to cases of sporadic ALS (sALS; n = 4) and familial ALS without known mutations (fALS; n = 2). We performed colorimetric immunohistochemistry (IHC) using antibodies against TDP-43, FUS/TLS, SOD1, C9orf72, ubiquitin, sequestosome 1 (p62), optineurin, phosphorylated high molecular weight neurofilament, peripherin, and Rho-guanine nucleotide exchange factor (RGNEF). We observed that RGNEF-immunoreactive neuronal cytoplasmic inclusions (NCIs) can co-localize with TDP-43, FUS/TLS and p62 within spinal MNs. We confirmed their capacity to interact by co-immunoprecipitations. We also found that mtSOD1-ALS cases possess a unique IHC signature, including the presence of C9orf72-immunoreactive diffuse NCIs, which allows them to be distinguished from other variants of ALS at the level of light microscopy. These findings support the hypothesis that alterations in RNA metabolism are a core pathogenic pathway in ALS. We also conclude that routine IHC-based analysis of spinal MNs may aid in the identification of families not previously suspected to harbor SOD1 mutations.

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Acknowledgments

This work was supported by the McFeat Family Fund and the Michael Halls Endowment. The authors declare no conflicts of interest. Postmortem spinal cord tissues were provided by the Department of Pathology at the London Health Sciences Center at the University of Western Ontario, London, Canada, and were obtained using standard autopsy consents. We thank Drs Janice Robertson and John Ravits for making available additional mtSOD1 slides. We are thankful to the donors and their families who contributed to this work through the donation of tissues and their consent to diagnostic confirmation and research. We also wish to thank Dr. Stephen Pasternak for his assistance with Bitplane’s Imaris image analysis software.

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Authors and Affiliations

  1. Department of Pathology, Western University, 1151 Richmond Street, London, ON, N6A 5K8, Canada

    Brian A. Keller & Lee Cyn Ang

  2. Molecular Brain Research Group, Robarts Research Institute, P.O. Box 5015, 100 Perth Drive, London, ON, N6A 5K8, Canada

    Brian A. Keller, Kathryn Volkening, Cristian A. Droppelmann & Michael J. Strong

  3. Department of Clinical Neurological Sciences, Western University, 1151 Richmond Street, London, ON, N6A 5K8, Canada

    Kathryn Volkening & Michael J. Strong

  4. Department of Neuroscience, Mayo Clinic College of Medicine, 4500 San Pablo Road, Jacksonville, FL, 32224, USA

    Rosa Rademakers

  5. Rm. C7-120, UH-LHSC, 339 Windermere Road, London, ON, N6A 5A5, Canada

    Michael J. Strong

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  1. Brian A. Keller
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  2. Kathryn Volkening
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Correspondence to Michael J. Strong.

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Keller, B.A., Volkening, K., Droppelmann, C.A. et al. Co-aggregation of RNA binding proteins in ALS spinal motor neurons: evidence of a common pathogenic mechanism. Acta Neuropathol 124, 733–747 (2012). https://doi.org/10.1007/s00401-012-1035-z

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