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Cell-produced ?-synuclein is secreted in a calcium-dependent manner by exosomes and impacts neuronal survival

Cell-produced ?-synuclein is secreted in a calcium-dependent manner by exosomes and impacts neuronal survival
Cell-produced ?-synuclein is secreted in a calcium-dependent manner by exosomes and impacts neuronal survival
?-Synuclein is central in Parkinson's disease pathogenesis. Although initially ?-synuclein was considered a purely intracellular protein, recent data suggest that it can be detected in the plasma and CSF of humans and in the culture media of neuronal cells. To address a role of secreted ?-synuclein in neuronal homeostasis, we have generated wild-type ?-synuclein and ?-galactosidase inducible SH-SY5Y cells. Soluble oligomeric and monomeric species of ?-synuclein are readily detected in the conditioned media (CM) of these cells at concentrations similar to those observed in human CSF. We have found that, in this model, ?-synuclein is secreted by externalized vesicles in a calcium-dependent manner. Electron microscopy and liquid chromatography–mass spectrometry proteomic analysis demonstrate that these vesicles have the characteristic hallmarks of exosomes, secreted intraluminar vesicles of multivesicular bodies. Application of CM containing secreted ?-synuclein causes cell death of recipient neuronal cells, which can be reversed after ?-synuclein immunodepletion from the CM. High- and low-molecular-weight ?-synuclein species, isolated from this CM, significantly decrease cell viability. Importantly, treatment of the CM with oligomer-interfering compounds before application rescues the recipient neuronal cells from the observed toxicity. Our results show for the first time that cell-produced ?-synuclein is secreted via an exosomal, calcium-dependent mechanism and suggest that ?-synuclein secretion serves to amplify and propagate Parkinson's disease-related pathology.

6838-6351
Emmanouilidou, Evangelia
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Melachroinou, Katerina
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Roumeliotis, Theodoros
f1284c98-b5eb-483e-9416-594c678e62fd
Garbis, Spiros D.
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Ntzouni, Maria
ee6db0b9-dc67-42f8-bd77-8ee3282c8f8c
Margaritis, Lukas H.
a5ee1423-d9a9-4c68-b0ff-2d81ca59c65c
Stefanis, Leonidas
a6f1880a-e589-4ffc-b274-dbc409f3e71d
Vekrellis, Kostas
0d76d598-854b-4dd6-9db1-7603580f4bc2
Emmanouilidou, Evangelia
e27a9aaf-4069-4e09-93a4-a3690710650f
Melachroinou, Katerina
c6a64834-f97c-4fca-b0dc-87c8abc0129f
Roumeliotis, Theodoros
f1284c98-b5eb-483e-9416-594c678e62fd
Garbis, Spiros D.
7067fd19-50c9-4d42-9611-f370289470bd
Ntzouni, Maria
ee6db0b9-dc67-42f8-bd77-8ee3282c8f8c
Margaritis, Lukas H.
a5ee1423-d9a9-4c68-b0ff-2d81ca59c65c
Stefanis, Leonidas
a6f1880a-e589-4ffc-b274-dbc409f3e71d
Vekrellis, Kostas
0d76d598-854b-4dd6-9db1-7603580f4bc2

Emmanouilidou, Evangelia, Melachroinou, Katerina, Roumeliotis, Theodoros, Garbis, Spiros D., Ntzouni, Maria, Margaritis, Lukas H., Stefanis, Leonidas and Vekrellis, Kostas (2010) Cell-produced ?-synuclein is secreted in a calcium-dependent manner by exosomes and impacts neuronal survival. Journal of Neuroscience, 30 (20), 6838-6351. (doi:10.1523/JNEUROSCI.5699-09.2010). (PMID:20484626)

Record type: Article

Abstract

?-Synuclein is central in Parkinson's disease pathogenesis. Although initially ?-synuclein was considered a purely intracellular protein, recent data suggest that it can be detected in the plasma and CSF of humans and in the culture media of neuronal cells. To address a role of secreted ?-synuclein in neuronal homeostasis, we have generated wild-type ?-synuclein and ?-galactosidase inducible SH-SY5Y cells. Soluble oligomeric and monomeric species of ?-synuclein are readily detected in the conditioned media (CM) of these cells at concentrations similar to those observed in human CSF. We have found that, in this model, ?-synuclein is secreted by externalized vesicles in a calcium-dependent manner. Electron microscopy and liquid chromatography–mass spectrometry proteomic analysis demonstrate that these vesicles have the characteristic hallmarks of exosomes, secreted intraluminar vesicles of multivesicular bodies. Application of CM containing secreted ?-synuclein causes cell death of recipient neuronal cells, which can be reversed after ?-synuclein immunodepletion from the CM. High- and low-molecular-weight ?-synuclein species, isolated from this CM, significantly decrease cell viability. Importantly, treatment of the CM with oligomer-interfering compounds before application rescues the recipient neuronal cells from the observed toxicity. Our results show for the first time that cell-produced ?-synuclein is secreted via an exosomal, calcium-dependent mechanism and suggest that ?-synuclein secretion serves to amplify and propagate Parkinson's disease-related pathology.

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Published date: 19 May 2010
Organisations: Cancer Sciences

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Local EPrints ID: 339292
URI: http://eprints.soton.ac.uk/id/eprint/339292
PURE UUID: be406156-4c09-4860-81b4-c46236fec628
ORCID for Spiros D. Garbis: ORCID iD orcid.org/0000-0002-1050-0805

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Date deposited: 28 May 2012 13:48
Last modified: 14 Mar 2024 11:13

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Contributors

Author: Evangelia Emmanouilidou
Author: Katerina Melachroinou
Author: Theodoros Roumeliotis
Author: Spiros D. Garbis ORCID iD
Author: Maria Ntzouni
Author: Lukas H. Margaritis
Author: Leonidas Stefanis
Author: Kostas Vekrellis

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