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Betemps D.,ANSES French Agency for Food | Verchere J.,ANSES French Agency for Food | Mougenot A.-L.,Indicia Production | Lachmann I.,AJ Roboscreen GmbH | And 5 more authors.
Journal of Visualized Experiments | Year: 2015

In addition to established methods like Western blot, new methods are needed to quickly and easily quantify disease-associated α-synuclein (αSD) 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 αSD (disease-associated αS) in these mice, an ELISA was designed to specifically quantify αSD in sick mice. Analysis of the central nervous system in this mouse model showed the presence of αSD mainly in the caudal brain regions and the spinal cord. There were no differences in αSD 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 αSD 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 Cterminal part of αS. The propagation of αSD, suggesting the involvement of a “prion-like” mechanism, can thus be easily monitored and quantified in this mouse model using an ELISA approach. © 2015 Journal of Visualized Experiments. Source

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