Detection of Disease-associated α-synuclein by Enhanced ELISA in the Brain of Transgenic Mice Overexpressing Human A53T Mutated α-synuclein
| Autor: | Latifa Lakhdar, Anne-Laure Mougenot, Dominique Bétemps, Stéphane Legastelois, Emilie Antier, Eric Morignat, Thierry Baron, Ingolf Lachmann, Jérémy Verchère |
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| Rok vydání: | 2015 |
| Předmět: |
Alpha-synuclein
Genetically modified mouse biology medicine.diagnostic_test General Immunology and Microbiology business.industry Transgene General Chemical Engineering General Neuroscience Central nervous system Molecular biology General Biochemistry Genetics and Molecular Biology Blot chemistry.chemical_compound medicine.anatomical_structure chemistry Western blot biology.protein medicine Immunohistochemistry Antibody business |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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