A cell culture model for monitoring α-synuclein cell-to-cell transfer.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4581567
- author
- Reyes, Juan F LU ; Olsson, Tomas LU ; Lamberts, Jennifer T ; Devine, Michael J ; Kunath, Tilo and Brundin, Patrik LU
- organization
- publishing date
- 2015
- 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
- 2023-01-02 06:58:11
@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}},
}