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Evaluation of α-synuclein immunohistochemical methods used by invited experts

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Abstract

The use of α-synuclein immunohistochemistry has altered our concepts of the cellular pathology, anatomical distribution and prevalence of Lewy body disorders. However, the diversity of methodology between laboratories has led to some inconsistencies in the literature. Adoption of uniformly sensitive methods may resolve some of these differences. Eight different immunohistochemical methods for demonstrating α-synuclein pathology, developed in eight separate expert laboratories, were evaluated for their sensitivity for neuronal elements affected by human Lewy body disorders. Identical test sets of formalin-fixed, paraffin-embedded sections from subjects diagnosed neuropathologically with or without Lewy body disorders were stained with the eight methods and graded by three observers for specific and nonspecific staining. The methods did not differ significantly in terms of Lewy body counts, but varied considerably in their ability to reveal neuropil elements such as fibers and dots. One method was clearly superior for revealing these neuropil elements and the critical factor contributing to its high sensitivity was considered to be its use of proteinase K as an epitope retrieval method. Some methods, however, achieved relatively high sensitivities with optimized formic acid protocols combined with a hydrolytic step. One method was developed that allows high sensitivity with commercially available reagents.

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Acknowledgments

The Sun Health Research Institute Brain Donation Program is supported by the National Institute on Aging (P30 AG19610 Arizona Alzheimer’s Disease Core Center), the Arizona Department of Health Services (contract 211002, Arizona Alzheimer’s Research Center), the Arizona Biomedical Research Commission (contracts 4001, 0011 and 05-901 to the Arizona Parkinson’s Disease Consortium) and the Prescott Family Initiative of the Michael J. Fox Foundation for Parkinson’s Research. Dr. Roncaroli is supported by the United Kingdom Parkinson’s Disease Society Tissue Bank. Dr. Leverenz is supported by the Department of Veterans Affairs.

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Authors and Affiliations

  1. Civin Laboratory for Neuropathology, Sun Health Research Institute, 10515 West Santa Fe Drive, Sun City, AZ, 85351, USA

    Thomas G. Beach & Lucia I. Sue

  2. Neuropathology Laboratory and Pathology Immunohistochemistry Laboratory, University of Texas Southwestern Medical School, Dallas, TX, USA

    Charles L. White & Christa L. Hladik

  3. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Ronald L. Hamilton

  4. Parkinson’s Disease Research, Education and Clinical Center, Philadelphia Veterans Affairs Medical Center, Philadelphia, PA, USA

    John E. Duda & Joseph V. Noorigian

  5. Department of Life-Pharmaceutics, University of Tokyo, Tokyo, Japan

    Takeshi Iwatsubo

  6. Department of Laboratory Medicine and Pathology, Mayo Clinic, Jacksonville, FL, USA

    Dennis W. Dickson

  7. Department of Neurology, University of Washington, Seattle, WA, USA

    James B. Leverenz

  8. Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, WA, USA

    James B. Leverenz

  9. Veterans Affairs Puget Sound Health Care System, Seattle, WA, USA

    James B. Leverenz

  10. Department of Clinical Neuroscience, Imperial College, London, UK

    Federico Roncaroli

  11. Elan Pharmaceuticals, San Francisco, CA, USA

    Manuel Buttini

  12. Department of Neurology, Mayo Clinic, Scottsdale, AZ, USA

    Charles H. Adler

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  1. Thomas G. Beach
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Correspondence to Thomas G. Beach.

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Beach, T.G., White, C.L., Hamilton, R.L. et al. Evaluation of α-synuclein immunohistochemical methods used by invited experts. Acta Neuropathol 116, 277–288 (2008). https://doi.org/10.1007/s00401-008-0409-8

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  • DOI: https://doi.org/10.1007/s00401-008-0409-8

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