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A cell culture model for monitoring α-synuclein cell-to-cell transfer.

Reyes, Juan F LU ; Olsson, Tomas LU ; Lamberts, Jennifer T ; Devine, Michael J ; Kunath, Tilo and Brundin, Patrik LU (2015) In Neurobiology of Disease 77(Jul 16). p.266-275
Abstract
The transfer of α-synuclein (α-syn) between cells has been proposed to be the primary mechanism of disease spreading in Parkinson's disease. Several cellular models exist that monitor the uptake of recombinant α-syn from the culture medium. Here we established a more physiologically relevant model system in which α-syn is produced and transferred between mammalian neurons. We generated cell lines expressing either α-syn tagged with fluorescent proteins or fluorescent tags alone then we co-cultured these cell lines to measure protein uptake. We used live-cell imaging to demonstrate intercellular α-syn transfer and used flow cytometry and high content analysis to quantify the transfer. We then successfully inhibited intercellular protein... (More)
The transfer of α-synuclein (α-syn) between cells has been proposed to be the primary mechanism of disease spreading in Parkinson's disease. Several cellular models exist that monitor the uptake of recombinant α-syn from the culture medium. Here we established a more physiologically relevant model system in which α-syn is produced and transferred between mammalian neurons. We generated cell lines expressing either α-syn tagged with fluorescent proteins or fluorescent tags alone then we co-cultured these cell lines to measure protein uptake. We used live-cell imaging to demonstrate intercellular α-syn transfer and used flow cytometry and high content analysis to quantify the transfer. We then successfully inhibited intercellular protein transfer genetically by down-regulating dynamin or pharmacologically using dynasore or heparin. In addition, we differentiated human induced pluripotent stem cells carrying a triplication of the α-syn gene into dopaminergic neurons. These cells secreted high levels of α-syn, which was taken up by neighboring neurons. Collectively, our co-culture systems provide simple but physiologically relevant tools for the identification of genetic modifiers or small molecules that inhibit α-syn cell-to-cell transfer. (Less)
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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Neurobiology of Disease
volume
77
issue
Jul 16
pages
266 - 275
publisher
Elsevier
external identifiers
  • pmid:25046995
  • wos:000353612200024
  • scopus:84939884559
  • pmid:25046995
ISSN
0969-9961
DOI
10.1016/j.nbd.2014.07.003
language
English
LU publication?
yes
id
6f94b82b-6975-4947-b6ba-a8d080c5bc15 (old id 4581567)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/25046995?dopt=Abstract
date added to LUP
2016-04-01 10:41:52
date last changed
2022-05-06 00:26:14
@article{6f94b82b-6975-4947-b6ba-a8d080c5bc15,
  abstract     = {{The transfer of α-synuclein (α-syn) between cells has been proposed to be the primary mechanism of disease spreading in Parkinson's disease. Several cellular models exist that monitor the uptake of recombinant α-syn from the culture medium. Here we established a more physiologically relevant model system in which α-syn is produced and transferred between mammalian neurons. We generated cell lines expressing either α-syn tagged with fluorescent proteins or fluorescent tags alone then we co-cultured these cell lines to measure protein uptake. We used live-cell imaging to demonstrate intercellular α-syn transfer and used flow cytometry and high content analysis to quantify the transfer. We then successfully inhibited intercellular protein transfer genetically by down-regulating dynamin or pharmacologically using dynasore or heparin. In addition, we differentiated human induced pluripotent stem cells carrying a triplication of the α-syn gene into dopaminergic neurons. These cells secreted high levels of α-syn, which was taken up by neighboring neurons. Collectively, our co-culture systems provide simple but physiologically relevant tools for the identification of genetic modifiers or small molecules that inhibit α-syn cell-to-cell transfer.}},
  author       = {{Reyes, Juan F and Olsson, Tomas and Lamberts, Jennifer T and Devine, Michael J and Kunath, Tilo and Brundin, Patrik}},
  issn         = {{0969-9961}},
  language     = {{eng}},
  number       = {{Jul 16}},
  pages        = {{266--275}},
  publisher    = {{Elsevier}},
  series       = {{Neurobiology of Disease}},
  title        = {{A cell culture model for monitoring α-synuclein cell-to-cell transfer.}},
  url          = {{http://dx.doi.org/10.1016/j.nbd.2014.07.003}},
  doi          = {{10.1016/j.nbd.2014.07.003}},
  volume       = {{77}},
  year         = {{2015}},
}