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Lookup NU author(s): Professor Tiago OuteiroORCiD
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© 2022 Elsevier B.V. Extracellular vesicles (EVs) are important mediators in intercellular communication. However, understanding the biological origin and functional effects of EVs subtypes has been challenging due to the moderate differences in their physical properties and absence of reliable markers. Here, we characterize the proteomes of ectosomes and exosomes using an improved differential ultracentrifugation protocol and quantitative proteomics. Our analyses revealed singular proteomic profiles for ectosomes and exosomes that enabled us to establish specific protein markers that can be used for their biochemical distinction. Cytoskeleton and glycolytic proteins are distinctively present in ectosomes, while endosomal sorting complexes proteins and tetraspanins are enriched in exosomes. Furthermore, annexin-A2 was identified as a specific marker for ectosomes derived from cell media and human cerebrospinal fluid. Expression of EGFP as a cytosolic reporter leads to its incorporation in EVs and enables their imaging with higher resolution. Assessment of neuronal network activity using multi-electrode array recordings demonstrated that spontaneous neuronal activity can be modulated by EVs. Ectosomes and exosomes internalization in neuronal cells disrupted their regular synchronized bursting activity, resulting in overall lower and more disorganized spiking activity. Our findings suggest that EVs cargoes reflect core intracellular processes, and their functional properties might regulate basic biological and pathological processes. Significance: This article presents novel approaches for studying the origin, composition, and biological effects in neuronal activity of ectosomes and exosomes. Our findings suggest that EVs cargoes reflect core intracellular processes, and their functional properties might regulate basic biological and pathological processes. Ultimately, our study also forms the foundation for future biomarker studies and for the understanding of the molecular basis of different diseases.
Author(s): Bras IC, Khani MH, Riedel D, Parfentev I, Gerhardt E, van Riesen C, Urlaub H, Gollisch T, Outeiro TF
Publication type: Article
Publication status: Published
Journal: Journal of Proteomics
Print publication date: 30/10/2022
Online publication date: 08/09/2022
Acceptance date: 02/09/2022
ISSN (print): 1874-3919
ISSN (electronic): 1876-7737
Publisher: Elsevier BV
PubMed id: 36089191
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