TY - JOUR
T1 - Characterization of Plasma Membrane Ceramides by Super-Resolution Microscopy
AU - Burgert, Anne
AU - Schlegel, Jan
AU - Bécam, Jérôme
AU - Doose, Sören
AU - Bieberich, Erhard
AU - Schubert-Unkmeir, Alexandra
AU - Sauer, Markus
N1 - Funding Information:
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.).
Publisher Copyright:
© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2017/5/22
Y1 - 2017/5/22
N2 - 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.
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.
KW - ceramide-rich platforms
KW - ceramides
KW - dSTORM
KW - sphingolipids
KW - super-resolution microscopy
UR - http://www.scopus.com/inward/record.url?scp=85017283080&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85017283080&partnerID=8YFLogxK
U2 - 10.1002/anie.201700570
DO - 10.1002/anie.201700570
M3 - Article
AN - SCOPUS:85017283080
SN - 1433-7851
VL - 56
SP - 6131
EP - 6135
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 22
ER -