Characterization of Plasma Membrane Ceramides by Super-Resolution Microscopy

Anne Burgert; Jan Schlegel; Jérôme Bécam; Sören Doose; Erhard Bieberich; Alexandra Schubert-Unkmeir; Markus Sauer

doi:10.1002/anie.201700570

Characterization of Plasma Membrane Ceramides by Super-Resolution Microscopy

Anne Burgert, Jan Schlegel, Jérôme Bécam, Sören Doose, Erhard Bieberich, Alexandra Schubert-Unkmeir, Markus Sauer

Physiology

Research output: Contribution to journal › Article › peer-review

49 Scopus citations

Abstract

The sphingolipid ceramide regulates cellular processes such as differentiation, proliferation, growth arrest, and apoptosis. Ceramide-rich membrane areas promote structural changes within the plasma membrane that segregate membrane receptors and affect membrane curvature and vesicle formation, fusion, and trafficking. Ceramides were labeled by immunocytochemistry to visualize their distribution on the plasma membrane of different cells with virtually molecular resolution by direct stochastic optical reconstruction microscopy (dSTORM). Super-resolution images show that independent of labeling conditions and cell type 50–60 % of all membrane ceramides are located in ceramide-rich platforms (CRPs) with a size of about 75 nm that are composed of at least about 20 ceramides. Treatment of cells with Bacillus cereus sphingomyelinase (bSMase) increases the overall ceramide concentration in the plasma membrane, the quantity of CRPs, and their size. Simultaneously, the ceramide concentration in CRPs increases approximately twofold.

Original language	English
Pages (from-to)	6131-6135
Number of pages	5
Journal	Angewandte Chemie - International Edition
Volume	56
Issue number	22
DOIs	https://doi.org/10.1002/anie.201700570
State	Published - May 22 2017

Bibliographical note

Publisher Copyright:
© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Funding

We thank Lisa Pliess-Behringer and Petra Gessner for assistance in cell culture and Christian Franke for data simulation. This work was supported by the Deutsche Forschungsgemeinschaft (DFG) (SA829/13-1 to M.S. and SCHU2394/2-1 to A.S.-U.) and the NIH (R01AG034389-01A2, R56NS095215-01, and NSF1121579 to E.B.).

Funders	Funder number
National Institutes of Health (NIH)	R56NS095215-01, NSF1121579
National Institute on Aging	R01AG034389
Deutsche Forschungsgemeinschaft	SCHU2394/2-1, SA829/13-1

Keywords

ceramide-rich platforms
ceramides
dSTORM
sphingolipids
super-resolution microscopy

ASJC Scopus subject areas

Catalysis
General Chemistry

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10.1002/anie.201700570

Cite this

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abstract = "The sphingolipid ceramide regulates cellular processes such as differentiation, proliferation, growth arrest, and apoptosis. Ceramide-rich membrane areas promote structural changes within the plasma membrane that segregate membrane receptors and affect membrane curvature and vesicle formation, fusion, and trafficking. Ceramides were labeled by immunocytochemistry to visualize their distribution on the plasma membrane of different cells with virtually molecular resolution by direct stochastic optical reconstruction microscopy (dSTORM). Super-resolution images show that independent of labeling conditions and cell type 50–60 \% of all membrane ceramides are located in ceramide-rich platforms (CRPs) with a size of about 75 nm that are composed of at least about 20 ceramides. Treatment of cells with Bacillus cereus sphingomyelinase (bSMase) increases the overall ceramide concentration in the plasma membrane, the quantity of CRPs, and their size. Simultaneously, the ceramide concentration in CRPs increases approximately twofold.",

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AU - Schlegel, Jan

AU - Bécam, Jérôme

AU - Doose, Sören

AU - Bieberich, Erhard

AU - Schubert-Unkmeir, Alexandra

AU - Sauer, Markus

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AB - The sphingolipid ceramide regulates cellular processes such as differentiation, proliferation, growth arrest, and apoptosis. Ceramide-rich membrane areas promote structural changes within the plasma membrane that segregate membrane receptors and affect membrane curvature and vesicle formation, fusion, and trafficking. Ceramides were labeled by immunocytochemistry to visualize their distribution on the plasma membrane of different cells with virtually molecular resolution by direct stochastic optical reconstruction microscopy (dSTORM). Super-resolution images show that independent of labeling conditions and cell type 50–60 % of all membrane ceramides are located in ceramide-rich platforms (CRPs) with a size of about 75 nm that are composed of at least about 20 ceramides. Treatment of cells with Bacillus cereus sphingomyelinase (bSMase) increases the overall ceramide concentration in the plasma membrane, the quantity of CRPs, and their size. Simultaneously, the ceramide concentration in CRPs increases approximately twofold.

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Characterization of Plasma Membrane Ceramides by Super-Resolution Microscopy

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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