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Comprehensive analysis of proteins encapsulated in brain-derived extracellular vesicles (BDEVs) from mice subjected to acute stress.

ABSTRACT: Brain-derived extracellular vesicles (BDEVs) were isolated from serum from acutely stressed and control mice by a combination of ultracentrifugation and immunoprecipitation, and a comprehensive analysis of the proteins contained within was performed.

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Yuji Morimoto

PROVIDER: PXD052683 | JPOST Repository | Fri May 30 00:00:00 BST 2025

REPOSITORIES: jPOST

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Project description:Extracellular vesicles (EVs) are involved in a wide range of physiological and pathological processes by shuttling material out of and between cells. Tissue EVs may thus lend insights into disease mechanisms and also betray disease when released into easily accessed biological fluids. Since brain-derived EVs (bdEVs) and their cargo may serve as biomarkers of neurodegenerative diseases, we evaluated modifications to a published, rigorous protocol for separation of EVs from brain tissue and studied effects of processing variables on quantitative and qualitative outcomes. To this end, size exclusion chromatography (SEC) and sucrose density gradient ultracentrifugation were compared as final separation steps in protocols involving stepped ultracentrifugation. bdEVs were separated from brain tissues of human, macaque. Effects of post-mortem interval (PMI) before final bdEV separation were probed. MISEV2018-compliant EV characterization was performed, and both small RNA and protein profiling were done. We conclude that the modified, SEC-employing protocol achieves EV separation efficiency roughly similar to a protocol using gradient density ultracentrifugation, while decreasing operator time and, potentially, variability. The protocol appears to yield bdEVs of higher purity for human tissues compared with macaque, suggesting opportunities for optimization. The interval between death/tissue storage/processing and final bdEV separation can also affect bdEV populations and composition and should thus be recorded for rigorous reporting. Different populations of EVs obtained through the modified method reported herein display characteristic RNA and protein content that hint at biomarker potential. To conclude, this study finds that the automatable and increasingly employed technique of SEC can be applied to tissue EV separation, and also reveals more about the importance of species-specific and technical considerations when working with tissue EVs. These results are expected to enhance the use of bdEVs in revealing and understanding brain disease.

			Action	DRS
	PK21038-1_Sample_A.mzML	Mzml
	PK21038-1_Sample_A.raw	Raw
	PK21038-2_Sample_B.mzML	Mzml
	PK21038-2_Sample_B.raw	Raw
	PK21038_Result.xlsx	Xlsx

Dataset Information

Comprehensive analysis of proteins encapsulated in brain-derived extracellular vesicles (BDEVs) from mice subjected to acute stress.

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