A novel panel of α-synuclein antibodies reveal distinctive staining profiles in synucleinopathies

Abstract

Synucleinopathies are a spectrum of neurodegenerative diseases characterized by the intracellular deposition of the protein α-synuclein leading to multiple outcomes, including dementia and Parkinsonism. Recent findings support the notion that across the spectrum of synucleinopathies there exist diverse but specific biochemical modifications and/or structural conformations of α-synuclein, which would give rise to protein strain specific prion-like intercellular transmission, a proposed model that could explain synucleinopathies disease progression. Herein, we characterized a panel of antibodies with epitopes within both the C- and N- termini of α-synuclein. A comprehensive analysis of human pathological tissue and mouse models of synucleinopathy with these antibodies support the notion that α-synuclein exists in distinct modified forms and/or structural variants. Furthermore, these well-characterized and specific tools allow the investigation of biochemical changes associated with α-synuclein inclusion formation. We have identified several antibodies of interest with diverse staining and epitope properties that will prove useful in future investigations of strain specific disease progression and the development of targeted immunotherapeutic approaches to synucleinopathies.

Skin α-synuclein deposits differ in clinical variants of synucleinopathy: an in vivo study

“…Similar results achieved in all clinical variants underlined that our data are reproducible and the assumption is accurate; (2) no antibodies against N-terminus α-syn epitopes were used. A recent work showed that N-terminus antibodies efficiently differentiated misfolded α-syn deposits in MSA from other clinical variants supporting a different conformation of misfolded deposits 44 . A future in vivo study targeting N-terminal epitopes is needed to ascertain this important point; (3) the number of PAF patients recruited for this study is limited but the number of aggregates studied is similar to the other variants of synucleinopathy; 4) although we studied a high number of abnormal intraneural α-syn deposits investigated subjects were fewer.…”

Carboxy‐terminal truncations of mouse α‐synuclein alter aggregation and prion‐like seeding

“…Mouse monoclonal antibody 81A (RRID: AB_2819037) is mainly specific for phosphorylated Ser129 (pSer129) αsyn and is a common diagnostic marker of pathologic αsyn ; the antibody was used at a 1 : 3000 dilution for immunohistochemistry. Antibody 94‐3A10 (RRID: AB_2819044) is a mouse monoclonal antibody specific for an epitope located at the last 10 C‐terminal amino acid of αsyn (residues 130‐140 αsyn) ; the antibody was used at a 1 : 10 000 dilution for immunohistochemistry and 1 : 1000 for western blotting. SNL‐4 (RRID: AB_2819043) is a rabbit polyclonal antibody raised against residues 2‐12 of αsyn ; the antibody was used at a 1 : 1000 dilution for western blotting.…”

Physiological C-terminal truncation of α-synuclein potentiates the prion-like formation of pathological inclusions

“…To investigate the ability of fibrils composed entirely of C-truncated forms of ␣syn to template polymerization of FL ␣syn, HEK293T cells were transfected with the plasmid expressing FL ␣syn and treated with 1 M preformed fibrils individually composed of each of the eight truncations used in this study or the 21-140 ␣syn control. The antibody 94-3A10 that requires residues 135-140 in ␣syn does not detect any C-truncations used and was used to detect FL ␣syn expressed in the cells (80). Most of the fibrils composed solely of C-truncated forms of ␣syn seeded FL ␣syn less efficiently than 21-140 ␣syn, and 1-122 and 1-124 ␣syn fibrils induced little if any FL ␣syn aggregation (Fig.…”