Extracellular vesicles containing ceramide-rich platforms: “mobile raft” isolation and analysis

Ahmed Elsherbini; Haiyan Qin; Zhihui Zhu; Priyanka Tripathi; Guanghu Wang; Simone M. Crivelli; Stefka D. Spassieva; Erhard Bieberich

doi:10.1007/978-1-0716-0814-2_5

Extracellular vesicles containing ceramide-rich platforms: “mobile raft” isolation and analysis

Ahmed Elsherbini, Haiyan Qin, Zhihui Zhu, Priyanka Tripathi, Guanghu Wang, Simone M. Crivelli, Stefka D. Spassieva, Erhard Bieberich

Physiology

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

6 Scopus citations

Abstract

Extracellular vesicles (EVs) are secreted by eukaryotic cells and serve as carriers for a variety of cell signaling factors, including RNAs, proteins, and lipids. We described a unique population of EVs, the membrane of which is highly enriched with the sphingolipid ceramide. We suggested that ceramide in the EV membrane is organized in ceramide-rich platforms (CRPs), a type of lipid raft that mediates interaction of ceramide with ceramide-associated proteins (CAPs). Here, we describe methods using anti-ceramide antibody to isolate ceramide-enriched EVs and detect exosomes after uptake into recipient cells. In addition, we discuss methods for EV analysis using nanoparticle tracking and mass spectrometry. The methods can be extended to the isolation of other types of EVs and “mobile rafts” transported by EVs from donor to recipient cells using antibodies against lipids specific for these EVs.

Original language	English
Title of host publication	Methods in Molecular Biology
Pages	87-98
Number of pages	12
DOIs	https://doi.org/10.1007/978-1-0716-0814-2_5
State	Published - 2021

Publication series

Name	Methods in Molecular Biology
Volume	2187
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Funding Information:
This work was in part supported by NIH grants R01AG034389 and R01AG064234, and Department of Veteran Affairs grant I01 BX003643. The authors are grateful for support by the Department of Physiology (Chair Dr. Alan Daugherty) at the University of Kentucky, Lexington, KY.

Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Affinity chromatography
Alzheimer’s disease
Ceramide-rich platform
Extracellular vesicles
Sphingolipid

ASJC Scopus subject areas

Molecular Biology
Genetics

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10.1007/978-1-0716-0814-2_5

Cite this

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abstract = "Extracellular vesicles (EVs) are secreted by eukaryotic cells and serve as carriers for a variety of cell signaling factors, including RNAs, proteins, and lipids. We described a unique population of EVs, the membrane of which is highly enriched with the sphingolipid ceramide. We suggested that ceramide in the EV membrane is organized in ceramide-rich platforms (CRPs), a type of lipid raft that mediates interaction of ceramide with ceramide-associated proteins (CAPs). Here, we describe methods using anti-ceramide antibody to isolate ceramide-enriched EVs and detect exosomes after uptake into recipient cells. In addition, we discuss methods for EV analysis using nanoparticle tracking and mass spectrometry. The methods can be extended to the isolation of other types of EVs and “mobile rafts” transported by EVs from donor to recipient cells using antibodies against lipids specific for these EVs.",

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note = "Funding Information: This work was in part supported by NIH grants R01AG034389 and R01AG064234, and Department of Veteran Affairs grant I01 BX003643. The authors are grateful for support by the Department of Physiology (Chair Dr. Alan Daugherty) at the University of Kentucky, Lexington, KY. Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",

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AU - Zhu, Zhihui

AU - Tripathi, Priyanka

AU - Wang, Guanghu

AU - Crivelli, Simone M.

AU - Spassieva, Stefka D.

AU - Bieberich, Erhard

N1 - Funding Information: This work was in part supported by NIH grants R01AG034389 and R01AG064234, and Department of Veteran Affairs grant I01 BX003643. The authors are grateful for support by the Department of Physiology (Chair Dr. Alan Daugherty) at the University of Kentucky, Lexington, KY. Publisher Copyright: © 2021, Springer Science+Business Media, LLC, part of Springer Nature.

PY - 2021

Y1 - 2021

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AB - Extracellular vesicles (EVs) are secreted by eukaryotic cells and serve as carriers for a variety of cell signaling factors, including RNAs, proteins, and lipids. We described a unique population of EVs, the membrane of which is highly enriched with the sphingolipid ceramide. We suggested that ceramide in the EV membrane is organized in ceramide-rich platforms (CRPs), a type of lipid raft that mediates interaction of ceramide with ceramide-associated proteins (CAPs). Here, we describe methods using anti-ceramide antibody to isolate ceramide-enriched EVs and detect exosomes after uptake into recipient cells. In addition, we discuss methods for EV analysis using nanoparticle tracking and mass spectrometry. The methods can be extended to the isolation of other types of EVs and “mobile rafts” transported by EVs from donor to recipient cells using antibodies against lipids specific for these EVs.

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KW - Ceramide-rich platform

KW - Extracellular vesicles

KW - Sphingolipid

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Extracellular vesicles containing ceramide-rich platforms: “mobile raft” isolation and analysis

Abstract

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Keywords

ASJC Scopus subject areas

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