title: Evaluation of alpha-synuclein immunohistochemical methods used by invited experts.
abstract: The use of alpha-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 alpha-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.
authors.affiliation: Civin Laboratory for Neuropathology, Sun Health Research Institute, 10515 West Santa Fe Drive, Sun City, AZ 85351, USA. thomas.beach@sunhealth.org
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Beach
authors.forename: Charles L
authors.initials: CL
authors.lastname: White
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Hamilton
authors.forename: John E
authors.initials: JE
authors.lastname: Duda
authors.forename: Takeshi
authors.initials: T
authors.lastname: Iwatsubo
authors.forename: Dennis W
authors.initials: DW
authors.lastname: Dickson
authors.forename: James B
authors.initials: JB
authors.lastname: Leverenz
authors.forename: Federico
authors.initials: F
authors.lastname: Roncaroli
authors.forename: Manuel
authors.initials: M
authors.lastname: Buttini
authors.forename: Christa L
authors.initials: CL
authors.lastname: Hladik
authors.forename: Lucia I
authors.initials: LI
authors.lastname: Sue
authors.forename: Joseph V
authors.initials: JV
authors.lastname: Noorigian
authors.forename: Charles H
authors.initials: CH
authors.lastname: Adler
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D020961:Lewy Body Disease / Q000378:metabolism* / Q000473:pathology; D016612:Paraffin Embedding; D010338:Pathology, Clinical / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D016707:Tissue Fixation; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein; D005557:Formaldehyde