title: Detection of Disease-associated α-synuclein by Enhanced ELISA in the Brain of Transgenic Mice Overexpressing Human A53T Mutated α-synuclein.
abstract: In addition to established methods like Western blot, new methods are needed to quickly and easily quantify disease-associated α-synuclein (αS(D)) in experimental models of synucleopathies. A transgenic mouse line (M83) over-expressing the human A53T αS and spontaneously developing a dramatic clinical phenotype between eight and 22 months of age, characterized by symptoms including weight loss, prostration, and severe motor impairment, was used in this study. For molecular analyses of αS(D) (disease-associated αS) in these mice, an ELISA was designed to specifically quantify αS(D) in sick mice. Analysis of the central nervous system in this mouse model showed the presence of αS(D) mainly in the caudal brain regions and the spinal cord. There were no differences in αS(D) distribution between different experimental conditions leading to clinical disease, i.e., in uninoculated and normally aging transgenic mice and in mice inoculated with brain extracts from sick mice. The specific detection of αS(D) immunoreactivity using an antibody against Ser129 phosphorylated αS by ELISA essentially correlated with that obtained by Western blot and immunohistochemistry. Unexpectedly, similar results were observed with several other antibodies against the C-terminal part of αS. The propagation of αS(D), suggesting the involvement of a "prion-like" mechanism, can thus be easily monitored and quantified in this mouse model using an ELISA approach.
authors.affiliation: Neurodegenerative Diseases Unit, ANSES - French Agency for Food, Environmental and Occupational Health & Safety.
authors.forename: Dominique
authors.initials: D
authors.lastname: Bétemps
authors.affiliation: Neurodegenerative Diseases Unit, ANSES - French Agency for Food, Environmental and Occupational Health & Safety.
authors.forename: Jérémy
authors.initials: J
authors.lastname: Verchère
authors.affiliation: Indicia Production.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Mougenot
authors.affiliation: AJ Roboscreen GmbH.
authors.forename: Ingolf
authors.initials: I
authors.lastname: Lachmann
authors.affiliation: Epidemiology Unit, ANSES - French Agency for Food, Environmental and Occupational Health & Safety.
authors.forename: Eric
authors.initials: E
authors.lastname: Morignat
authors.affiliation: Experimental Facilities, ANSES - French Agency for Food, Environmental and Occupational Health & Safety.
authors.forename: Emilie
authors.initials: E
authors.lastname: Antier
authors.affiliation: Experimental Facilities, ANSES - French Agency for Food, Environmental and Occupational Health & Safety.
authors.forename: Latifa
authors.initials: L
authors.lastname: Lakhdar
authors.affiliation: Indicia Production.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Legastelois
authors.affiliation: Neurodegenerative Diseases Unit, ANSES - French Agency for Food, Environmental and Occupational Health & Safety; Thierry.BARON@anses.fr.
authors.forename: Thierry
authors.initials: T
authors.lastname: Baron
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic; D010766:Phosphorylation; D011328:Prions / Q000378:metabolism; D013116:Spinal Cord / Q000378:metabolism; D051844:alpha-Synuclein / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011328:Prions; D051844:alpha-Synuclein