Single cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation
Abstract The intercellular spreading of protein assemblies is a major factor in the progression of neurodegenerative disorders. The quantitative study and visualization of cell-to-cell propagation using tagged-proteins is challenging due to the steric effect of relatively large fluorescence tags and...
Guardado en:
| Autores principales: | , , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Nature Portfolio
2017
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/d049f33829294b88ae864baa009409c9 |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:d049f33829294b88ae864baa009409c9 |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:d049f33829294b88ae864baa009409c92021-12-02T15:18:52ZSingle cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation10.1038/s41598-017-00657-z2045-2322https://doaj.org/article/d049f33829294b88ae864baa009409c92017-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00657-zhttps://doaj.org/toc/2045-2322Abstract The intercellular spreading of protein assemblies is a major factor in the progression of neurodegenerative disorders. The quantitative study and visualization of cell-to-cell propagation using tagged-proteins is challenging due to the steric effect of relatively large fluorescence tags and the risk of ‘false positive’ identification when analyzing these rare transmission events. Here, we established a cell culture model to characterize the cell-to-cell transmission of TAR DNA-binding protein and α-synuclein, involved in amyotrophic lateral sclerosis and Parkinson’s disease, respectively, using the small nine amino acid influenza hemagglutinin tag. The novel use of single cell resolution imaging flow cytometry allowed the visualization and quantification of all individual transmission events. Cell-level analysis of these events indicated that the degree of transfer is lower than previously reported based on conventional flow cytometry. Furthermore, our analysis can exclude ‘false positive’ events of cellular overlap and extracellular debris attachment. The results were corroborated by high-resolution confocal microscopy mapping of protein localization.Sivan PeledDorin SadeYaron BramZiv PoratTopaz KreiserMichael MimouniAlexandra LichtensteinDaniel SegalEhud GazitNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Sivan Peled Dorin Sade Yaron Bram Ziv Porat Topaz Kreiser Michael Mimouni Alexandra Lichtenstein Daniel Segal Ehud Gazit Single cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation |
| description |
Abstract The intercellular spreading of protein assemblies is a major factor in the progression of neurodegenerative disorders. The quantitative study and visualization of cell-to-cell propagation using tagged-proteins is challenging due to the steric effect of relatively large fluorescence tags and the risk of ‘false positive’ identification when analyzing these rare transmission events. Here, we established a cell culture model to characterize the cell-to-cell transmission of TAR DNA-binding protein and α-synuclein, involved in amyotrophic lateral sclerosis and Parkinson’s disease, respectively, using the small nine amino acid influenza hemagglutinin tag. The novel use of single cell resolution imaging flow cytometry allowed the visualization and quantification of all individual transmission events. Cell-level analysis of these events indicated that the degree of transfer is lower than previously reported based on conventional flow cytometry. Furthermore, our analysis can exclude ‘false positive’ events of cellular overlap and extracellular debris attachment. The results were corroborated by high-resolution confocal microscopy mapping of protein localization. |
| format |
article |
| author |
Sivan Peled Dorin Sade Yaron Bram Ziv Porat Topaz Kreiser Michael Mimouni Alexandra Lichtenstein Daniel Segal Ehud Gazit |
| author_facet |
Sivan Peled Dorin Sade Yaron Bram Ziv Porat Topaz Kreiser Michael Mimouni Alexandra Lichtenstein Daniel Segal Ehud Gazit |
| author_sort |
Sivan Peled |
| title |
Single cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation |
| title_short |
Single cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation |
| title_full |
Single cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation |
| title_fullStr |
Single cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation |
| title_full_unstemmed |
Single cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation |
| title_sort |
single cell imaging and quantification of tdp-43 and α-synuclein intercellular propagation |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/d049f33829294b88ae864baa009409c9 |
| work_keys_str_mv |
AT sivanpeled singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT dorinsade singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT yaronbram singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT zivporat singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT topazkreiser singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT michaelmimouni singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT alexandralichtenstein singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT danielsegal singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation AT ehudgazit singlecellimagingandquantificationoftdp43andasynucleinintercellularpropagation |
| _version_ |
1718387476322582528 |