Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain
Abstract Assays for quantifying aggregated and phosphorylated (S129) human α-synuclein protein are widely used to evaluate pathological burden in patients suffering from synucleinopathy disorders. Many of these assays, however, do not cross-react with mouse α-synuclein or exhibit poor sensitivity fo...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-11-01
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| Series: | npj Parkinson's Disease |
| Online Access: | https://doi.org/10.1038/s41531-024-00830-y |
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| author | Benjamin Guy Trist Courtney Jade Wright Alejandra Rangel Louise Cottle Asheeta Prasad Nanna Møller Jensen Hjalte Gram Nicolas Dzamko Poul Henning Jensen Deniz Kirik |
| author_facet | Benjamin Guy Trist Courtney Jade Wright Alejandra Rangel Louise Cottle Asheeta Prasad Nanna Møller Jensen Hjalte Gram Nicolas Dzamko Poul Henning Jensen Deniz Kirik |
| author_sort | Benjamin Guy Trist |
| collection | DOAJ |
| description | Abstract Assays for quantifying aggregated and phosphorylated (S129) human α-synuclein protein are widely used to evaluate pathological burden in patients suffering from synucleinopathy disorders. Many of these assays, however, do not cross-react with mouse α-synuclein or exhibit poor sensitivity for this target, which is problematic considering the preponderance of mouse models at the forefront of pre-clinical α-synuclein research. In this project, we addressed this unmet need by reformulating two existing AlphaLISA® SureFire® Ultra™ total and pS129 α-synuclein assay kits to yield robust and ultrasensitive (LLoQ ≤ 0.5 pg/mL) quantification of mouse and human wild-type and pS129 α-synuclein protein. We then employed these assays, together with the BioLegend α-synuclein aggregate ELISA, to assess α-synuclein S129 phosphorylation and aggregation in different mouse brain tissue preparations. Overall, we highlight the compatibility of these new immunoassays with rodent models and demonstrate their potential to advance knowledge surrounding α-synuclein phosphorylation and aggregation in synucleinopathies. |
| format | Article |
| id | doaj-art-f576a04dee31405487e4e7a89a22aae6 |
| institution | DOAJ |
| issn | 2373-8057 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Parkinson's Disease |
| spelling | doaj-art-f576a04dee31405487e4e7a89a22aae62025-08-20T02:49:56ZengNature Portfolionpj Parkinson's Disease2373-80572024-11-0110111410.1038/s41531-024-00830-yNovel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brainBenjamin Guy Trist0Courtney Jade Wright1Alejandra Rangel2Louise Cottle3Asheeta Prasad4Nanna Møller Jensen5Hjalte Gram6Nicolas Dzamko7Poul Henning Jensen8Deniz Kirik9Charles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of SydneyCharles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of SydneyCharles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of SydneyCharles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of SydneyCharles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of SydneyDepartment of Biomedicine, Aarhus UniversityDepartment of Biomedicine, Aarhus UniversityBrain and Mind Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of SydneyDepartment of Biomedicine, Aarhus UniversityCharles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of SydneyAbstract Assays for quantifying aggregated and phosphorylated (S129) human α-synuclein protein are widely used to evaluate pathological burden in patients suffering from synucleinopathy disorders. Many of these assays, however, do not cross-react with mouse α-synuclein or exhibit poor sensitivity for this target, which is problematic considering the preponderance of mouse models at the forefront of pre-clinical α-synuclein research. In this project, we addressed this unmet need by reformulating two existing AlphaLISA® SureFire® Ultra™ total and pS129 α-synuclein assay kits to yield robust and ultrasensitive (LLoQ ≤ 0.5 pg/mL) quantification of mouse and human wild-type and pS129 α-synuclein protein. We then employed these assays, together with the BioLegend α-synuclein aggregate ELISA, to assess α-synuclein S129 phosphorylation and aggregation in different mouse brain tissue preparations. Overall, we highlight the compatibility of these new immunoassays with rodent models and demonstrate their potential to advance knowledge surrounding α-synuclein phosphorylation and aggregation in synucleinopathies.https://doi.org/10.1038/s41531-024-00830-y |
| spellingShingle | Benjamin Guy Trist Courtney Jade Wright Alejandra Rangel Louise Cottle Asheeta Prasad Nanna Møller Jensen Hjalte Gram Nicolas Dzamko Poul Henning Jensen Deniz Kirik Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain npj Parkinson's Disease |
| title | Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain |
| title_full | Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain |
| title_fullStr | Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain |
| title_full_unstemmed | Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain |
| title_short | Novel tools to quantify total, phospho-Ser129 and aggregated alpha-synuclein in the mouse brain |
| title_sort | novel tools to quantify total phospho ser129 and aggregated alpha synuclein in the mouse brain |
| url | https://doi.org/10.1038/s41531-024-00830-y |
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