Detection Of Disease-Associated Alpha-Synuclein By Enhanced Elisa In The Brain Of Transgenic Mice Overexpressing Human A53t Mutated Alpha-Synuclein

In addition to established methods like Western blot, new methods are needed to quickly and easily quantify disease-associated alpha-synuclein (alpha S-D) in experimental models of synucleopathies. A transgenic mouse line (M83) over-expressing the human A53T alpha S 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 alpha S-D (disease-associated alpha S) in these mice, an ELISA was designed to specifically quantify alpha S-D in sick mice. Analysis of the central nervous system in this mouse model showed the presence of alpha S-D mainly in the caudal brain regions and the spinal cord. There were no differences in alpha S-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 alpha S-D immunoreactivity using an antibody against Ser129 phosphorylated alpha S 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 alpha S. The propagation of alpha S-D, suggesting the involvement of a "prion-like" mechanism, can thus be easily monitored and quantified in this mouse model using an ELISA approach.