Acta Neuropathologica

, 116:277

Evaluation of α-synuclein immunohistochemical methods used by invited experts

  • Thomas G. Beach
  • Charles L. White
  • Ronald L. Hamilton
  • John E. Duda
  • Takeshi Iwatsubo
  • Dennis W. Dickson
  • James B. Leverenz
  • Federico Roncaroli
  • Manuel Buttini
  • Christa L. Hladik
  • Lucia I. Sue
  • Joseph V. Noorigian
  • Charles H. Adler
Original Paper

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.

Supplementary material

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Thomas G. Beach
    • 1
  • Charles L. White
    • 2
  • Ronald L. Hamilton
    • 3
  • John E. Duda
    • 4
  • Takeshi Iwatsubo
    • 5
  • Dennis W. Dickson
    • 6
  • James B. Leverenz
    • 7
    • 8
    • 9
  • Federico Roncaroli
    • 10
  • Manuel Buttini
    • 11
  • Christa L. Hladik
    • 2
  • Lucia I. Sue
    • 1
  • Joseph V. Noorigian
    • 4
  • Charles H. Adler
    • 12
  1. 1.Civin Laboratory for NeuropathologySun Health Research InstituteSun CityAZUSA
  2. 2.Neuropathology Laboratory and Pathology Immunohistochemistry LaboratoryUniversity of Texas Southwestern Medical SchoolDallasUSA
  3. 3.Department of PathologyUniversity of Pittsburgh School of MedicinePittsburghUSA
  4. 4.Parkinson’s Disease Research, Education and Clinical CenterPhiladelphia Veterans Affairs Medical CenterPhiladelphiaUSA
  5. 5.Department of Life-PharmaceuticsUniversity of TokyoTokyoJapan
  6. 6.Department of Laboratory Medicine and PathologyMayo ClinicJacksonvilleUSA
  7. 7.Department of NeurologyUniversity of WashingtonSeattleUSA
  8. 8.Department of Psychiatry and Behavioral SciencesUniversity of WashingtonSeattleUSA
  9. 9.Veterans Affairs Puget Sound Health Care SystemSeattleUSA
  10. 10.Department of Clinical NeuroscienceImperial CollegeLondonUK
  11. 11.Elan PharmaceuticalsSan FranciscoUSA
  12. 12.Department of NeurologyMayo ClinicScottsdaleUSA

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