Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED Microscopy
α-Synuclein (αS) is an intrinsically disordered and highly dynamic protein involved in dopamine release at presynaptic terminals. The abnormal aggregation of αS as mature fibrils into intraneuronal inclusion bodies is directly linked to Parkinson’s disease. Increasing experimental evidence suggests...
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MDPI AG
2021-05-01
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| Series: | Life |
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| Online Access: | https://www.mdpi.com/2075-1729/11/5/431 |
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| author | Alessandra Bigi Emilio Ermini Serene W. Chen Roberta Cascella Cristina Cecchi |
| author_facet | Alessandra Bigi Emilio Ermini Serene W. Chen Roberta Cascella Cristina Cecchi |
| author_sort | Alessandra Bigi |
| collection | DOAJ |
| description | α-Synuclein (αS) is an intrinsically disordered and highly dynamic protein involved in dopamine release at presynaptic terminals. The abnormal aggregation of αS as mature fibrils into intraneuronal inclusion bodies is directly linked to Parkinson’s disease. Increasing experimental evidence suggests that soluble oligomers formed early during the aggregation process are the most cytotoxic forms of αS. This study investigated the uptake by neuronal cells of pathologically relevant αS oligomers and fibrils exploiting a range of conformation-sensitive antibodies, and the super-resolution stimulated emission depletion (STED) microscopy. We found that prefibrillar oligomers promptly penetrate neuronal membranes, thus resulting in cell dysfunction. By contrast, fibril docking to the phospholipid bilayer is accompanied by αS conformational changes with a progressive release of A11-reactive oligomers, which can enter into the neurons and trigger cell impairment. Our data provide important evidence on the role of αS fibrils as a source of harmful oligomers, which resemble the intermediate conformers formed de novo during aggregation, underling the dynamic and reversible nature of protein aggregates responsible for α-synucleinopathies. |
| first_indexed | 2024-03-10T11:32:53Z |
| format | Article |
| id | doaj.art-9780ceb0824c4c369a1076c296215bb5 |
| institution | Directory Open Access Journal |
| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-10T11:32:53Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Life |
| spelling | doaj.art-9780ceb0824c4c369a1076c296215bb52023-11-21T19:09:14ZengMDPI AGLife2075-17292021-05-0111543110.3390/life11050431Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED MicroscopyAlessandra Bigi0Emilio Ermini1Serene W. Chen2Roberta Cascella3Cristina Cecchi4Section of Biochemistry, Department of Experimental and Clinical Biomedical Sciences, University of Florence, 50134 Florence, ItalySection of Biochemistry, Department of Experimental and Clinical Biomedical Sciences, University of Florence, 50134 Florence, ItalyCentre for Misfolding Diseases, Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UKSection of Biochemistry, Department of Experimental and Clinical Biomedical Sciences, University of Florence, 50134 Florence, ItalySection of Biochemistry, Department of Experimental and Clinical Biomedical Sciences, University of Florence, 50134 Florence, Italyα-Synuclein (αS) is an intrinsically disordered and highly dynamic protein involved in dopamine release at presynaptic terminals. The abnormal aggregation of αS as mature fibrils into intraneuronal inclusion bodies is directly linked to Parkinson’s disease. Increasing experimental evidence suggests that soluble oligomers formed early during the aggregation process are the most cytotoxic forms of αS. This study investigated the uptake by neuronal cells of pathologically relevant αS oligomers and fibrils exploiting a range of conformation-sensitive antibodies, and the super-resolution stimulated emission depletion (STED) microscopy. We found that prefibrillar oligomers promptly penetrate neuronal membranes, thus resulting in cell dysfunction. By contrast, fibril docking to the phospholipid bilayer is accompanied by αS conformational changes with a progressive release of A11-reactive oligomers, which can enter into the neurons and trigger cell impairment. Our data provide important evidence on the role of αS fibrils as a source of harmful oligomers, which resemble the intermediate conformers formed de novo during aggregation, underling the dynamic and reversible nature of protein aggregates responsible for α-synucleinopathies.https://www.mdpi.com/2075-1729/11/5/431synucleinopathiesprotein aggregationamyloidtoxic oligomersLewy bodiesPD |
| spellingShingle | Alessandra Bigi Emilio Ermini Serene W. Chen Roberta Cascella Cristina Cecchi Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED Microscopy Life synucleinopathies protein aggregation amyloid toxic oligomers Lewy bodies PD |
| title | Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED Microscopy |
| title_full | Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED Microscopy |
| title_fullStr | Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED Microscopy |
| title_full_unstemmed | Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED Microscopy |
| title_short | Exploring the Release of Toxic Oligomers from α-Synuclein Fibrils with Antibodies and STED Microscopy |
| title_sort | exploring the release of toxic oligomers from α synuclein fibrils with antibodies and sted microscopy |
| topic | synucleinopathies protein aggregation amyloid toxic oligomers Lewy bodies PD |
| url | https://www.mdpi.com/2075-1729/11/5/431 |
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