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

Juan F. Reyes, Tomas T. Olsson, Jennifer T. Lamberts, Michael J. Devine, Tilo Kunath, Patrik Brundin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)266-275
Number of pages10
JournalNeurobiology of disease
Volume77
DOIs
Publication statusPublished - 12 Aug 2014

Keywords

  • Dynamin
  • Dynasore
  • Flow cytometry
  • HCA
  • Heparin
  • iPS cells
  • Parkinson's disease
  • Prion-like
  • Synucleinopathy

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