Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein
Abstract Background α-Synuclein (aSyn) aggregation is thought to play a central role in neurodegenerative disorders termed synucleinopathies, including Parkinson’s disease (PD). Mouse aSyn contains a threonine residue at position 53 that mimics the human familial PD substitution A53T, yet in contras...
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BMC
2020-09-01
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Series: | Molecular Neurodegeneration |
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Online Access: | http://link.springer.com/article/10.1186/s13024-020-00380-w |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Natalie Landeck Katherine E. Strathearn Daniel Ysselstein Kerstin Buck Sayan Dutta Siddhartha Banerjee Zhengjian Lv John D. Hulleman Jagadish Hindupur Li-Kai Lin Sonal Padalkar Lia A. Stanciu Yuri L. Lyubchenko Deniz Kirik Jean-Christophe Rochet |
spellingShingle |
Natalie Landeck Katherine E. Strathearn Daniel Ysselstein Kerstin Buck Sayan Dutta Siddhartha Banerjee Zhengjian Lv John D. Hulleman Jagadish Hindupur Li-Kai Lin Sonal Padalkar Lia A. Stanciu Yuri L. Lyubchenko Deniz Kirik Jean-Christophe Rochet Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein Molecular Neurodegeneration Adeno-associated virus Aggregation Alpha-synuclein Amyloid Fibril Membrane |
author_facet |
Natalie Landeck Katherine E. Strathearn Daniel Ysselstein Kerstin Buck Sayan Dutta Siddhartha Banerjee Zhengjian Lv John D. Hulleman Jagadish Hindupur Li-Kai Lin Sonal Padalkar Lia A. Stanciu Yuri L. Lyubchenko Deniz Kirik Jean-Christophe Rochet |
author_sort |
Natalie Landeck |
title |
Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein |
title_short |
Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein |
title_full |
Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein |
title_fullStr |
Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein |
title_full_unstemmed |
Two C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein |
title_sort |
two c-terminal sequence variations determine differential neurotoxicity between human and mouse α-synuclein |
publisher |
BMC |
series |
Molecular Neurodegeneration |
issn |
1750-1326 |
publishDate |
2020-09-01 |
description |
Abstract Background α-Synuclein (aSyn) aggregation is thought to play a central role in neurodegenerative disorders termed synucleinopathies, including Parkinson’s disease (PD). Mouse aSyn contains a threonine residue at position 53 that mimics the human familial PD substitution A53T, yet in contrast to A53T patients, mice show no evidence of aSyn neuropathology even after aging. Here, we studied the neurotoxicity of human A53T, mouse aSyn, and various human-mouse chimeras in cellular and in vivo models, as well as their biochemical properties relevant to aSyn pathobiology. Methods Primary midbrain cultures transduced with aSyn-encoding adenoviruses were analyzed immunocytochemically to determine relative dopaminergic neuron viability. Brain sections prepared from rats injected intranigrally with aSyn-encoding adeno-associated viruses were analyzed immunohistochemically to determine nigral dopaminergic neuron viability and striatal dopaminergic terminal density. Recombinant aSyn variants were characterized in terms of fibrillization rates by measuring thioflavin T fluorescence, fibril morphologies via electron microscopy and atomic force microscopy, and protein-lipid interactions by monitoring membrane-induced aSyn aggregation and aSyn-mediated vesicle disruption. Statistical tests consisted of ANOVA followed by Tukey’s multiple comparisons post hoc test and the Kruskal-Wallis test followed by a Dunn’s multiple comparisons test or a two-tailed Mann-Whitney test. Results Mouse aSyn was less neurotoxic than human aSyn A53T in cell culture and in rat midbrain, and data obtained for the chimeric variants indicated that the human-to-mouse substitutions D121G and N122S were at least partially responsible for this decrease in neurotoxicity. Human aSyn A53T and a chimeric variant with the human residues D and N at positions 121 and 122 (respectively) showed a greater propensity to undergo membrane-induced aggregation and to elicit vesicle disruption. Differences in neurotoxicity among the human, mouse, and chimeric aSyn variants correlated weakly with differences in fibrillization rate or fibril morphology. Conclusions Mouse aSyn is less neurotoxic than the human A53T variant as a result of inhibitory effects of two C-terminal amino acid substitutions on membrane-induced aSyn aggregation and aSyn-mediated vesicle permeabilization. Our findings highlight the importance of membrane-induced self-assembly in aSyn neurotoxicity and suggest that inhibiting this process by targeting the C-terminal domain could slow neurodegeneration in PD and other synucleinopathy disorders. |
topic |
Adeno-associated virus Aggregation Alpha-synuclein Amyloid Fibril Membrane |
url |
http://link.springer.com/article/10.1186/s13024-020-00380-w |
work_keys_str_mv |
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doaj-577ae13724214a3bac084228b03fe4b42020-11-25T03:31:02ZengBMCMolecular Neurodegeneration1750-13262020-09-0115112310.1186/s13024-020-00380-wTwo C-terminal sequence variations determine differential neurotoxicity between human and mouse α-synucleinNatalie Landeck0Katherine E. Strathearn1Daniel Ysselstein2Kerstin Buck3Sayan Dutta4Siddhartha Banerjee5Zhengjian Lv6John D. Hulleman7Jagadish Hindupur8Li-Kai Lin9Sonal Padalkar10Lia A. Stanciu11Yuri L. Lyubchenko12Deniz Kirik13Jean-Christophe Rochet14Brain Repair and Imaging in Neural Systems, Department of Experimental Medical Science, Lund UniversityDepartment of Medicinal Chemistry and Molecular Pharmacology, Purdue UniversityDepartment of Medicinal Chemistry and Molecular Pharmacology, Purdue UniversityBrain Repair and Imaging in Neural Systems, Department of Experimental Medical Science, Lund UniversityDepartment of Medicinal Chemistry and Molecular Pharmacology, Purdue UniversityDepartment of Pharmaceutical Sciences, University of Nebraska Medical CenterDepartment of Pharmaceutical Sciences, University of Nebraska Medical CenterDepartment of Medicinal Chemistry and Molecular Pharmacology, Purdue UniversityDepartment of Medicinal Chemistry and Molecular Pharmacology, Purdue UniversitySchool of Materials Engineering, Purdue UniversitySchool of Materials Engineering, Purdue UniversitySchool of Materials Engineering, Purdue UniversityDepartment of Pharmaceutical Sciences, University of Nebraska Medical CenterBrain Repair and Imaging in Neural Systems, Department of Experimental Medical Science, Lund UniversityDepartment of Medicinal Chemistry and Molecular Pharmacology, Purdue UniversityAbstract Background α-Synuclein (aSyn) aggregation is thought to play a central role in neurodegenerative disorders termed synucleinopathies, including Parkinson’s disease (PD). Mouse aSyn contains a threonine residue at position 53 that mimics the human familial PD substitution A53T, yet in contrast to A53T patients, mice show no evidence of aSyn neuropathology even after aging. Here, we studied the neurotoxicity of human A53T, mouse aSyn, and various human-mouse chimeras in cellular and in vivo models, as well as their biochemical properties relevant to aSyn pathobiology. Methods Primary midbrain cultures transduced with aSyn-encoding adenoviruses were analyzed immunocytochemically to determine relative dopaminergic neuron viability. Brain sections prepared from rats injected intranigrally with aSyn-encoding adeno-associated viruses were analyzed immunohistochemically to determine nigral dopaminergic neuron viability and striatal dopaminergic terminal density. Recombinant aSyn variants were characterized in terms of fibrillization rates by measuring thioflavin T fluorescence, fibril morphologies via electron microscopy and atomic force microscopy, and protein-lipid interactions by monitoring membrane-induced aSyn aggregation and aSyn-mediated vesicle disruption. Statistical tests consisted of ANOVA followed by Tukey’s multiple comparisons post hoc test and the Kruskal-Wallis test followed by a Dunn’s multiple comparisons test or a two-tailed Mann-Whitney test. Results Mouse aSyn was less neurotoxic than human aSyn A53T in cell culture and in rat midbrain, and data obtained for the chimeric variants indicated that the human-to-mouse substitutions D121G and N122S were at least partially responsible for this decrease in neurotoxicity. Human aSyn A53T and a chimeric variant with the human residues D and N at positions 121 and 122 (respectively) showed a greater propensity to undergo membrane-induced aggregation and to elicit vesicle disruption. Differences in neurotoxicity among the human, mouse, and chimeric aSyn variants correlated weakly with differences in fibrillization rate or fibril morphology. Conclusions Mouse aSyn is less neurotoxic than the human A53T variant as a result of inhibitory effects of two C-terminal amino acid substitutions on membrane-induced aSyn aggregation and aSyn-mediated vesicle permeabilization. Our findings highlight the importance of membrane-induced self-assembly in aSyn neurotoxicity and suggest that inhibiting this process by targeting the C-terminal domain could slow neurodegeneration in PD and other synucleinopathy disorders.http://link.springer.com/article/10.1186/s13024-020-00380-wAdeno-associated virusAggregationAlpha-synucleinAmyloidFibrilMembrane |