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Isolation and Characterization of Sympathetic Extracellular Vesicles

View ORCID ProfileAshley J Mason, Austin B Keeler, Farah Kabir, Bettina Winckler, Christopher Deppmann
doi: https://doi.org/10.1101/2022.05.24.493096
Ashley J Mason
1Department of Biology, University of Virginia, Charlottesville, Virginia, USA
2Department of Cell Biology, University of Virginia, Charlottesville, Virginia, USA
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  • ORCID record for Ashley J Mason
Austin B Keeler
1Department of Biology, University of Virginia, Charlottesville, Virginia, USA
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Farah Kabir
1Department of Biology, University of Virginia, Charlottesville, Virginia, USA
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Bettina Winckler
2Department of Cell Biology, University of Virginia, Charlottesville, Virginia, USA
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  • For correspondence: bw5h@virginia.edu deppmann@virginia.edu
Christopher Deppmann
1Department of Biology, University of Virginia, Charlottesville, Virginia, USA
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  • For correspondence: bw5h@virginia.edu deppmann@virginia.edu
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Abstract

Research on Extracellular Vesicles (EVs) is increasing at a rapid pace; therefore, it is crucial to maintain rigor in characterizing EVs from a new model system. Neuronal derived EVs have been well described in the central nervous system; however, studies in the peripheral nervous system have largely focused on EVs derived from supporting cell types such as endothelial cells or glia. Additionally, EVs are heterogeneous in size, shape, cargo and biogenic origin and therefore a multimodal approach to characterization must be used. Here we conduct a thorough description of EVs derived from sympathetic neurons using immunoblot assays, nanoparticle tracking analysis and cryo-electron microscopy. We show that primary sympathetic cultures secrete EVs in a density-dependent manner and that their sizing aligns with those reported in the literature. Lastly, using a compartmentalized culture system we show that EVs secreted by the somatodendritic domain of neurons contain cargo that originated at their distal axon. This work establishes foundational protocols to explore the biogenesis and function of EVs in the peripheral nervous system.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 25, 2022.
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Isolation and Characterization of Sympathetic Extracellular Vesicles
Ashley J Mason, Austin B Keeler, Farah Kabir, Bettina Winckler, Christopher Deppmann
bioRxiv 2022.05.24.493096; doi: https://doi.org/10.1101/2022.05.24.493096
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Isolation and Characterization of Sympathetic Extracellular Vesicles
Ashley J Mason, Austin B Keeler, Farah Kabir, Bettina Winckler, Christopher Deppmann
bioRxiv 2022.05.24.493096; doi: https://doi.org/10.1101/2022.05.24.493096

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