title: Characterization of Plasma Membrane Ceramides by Super-Resolution Microscopy.
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.
authors.affiliation: Department of Biotechnology and Biophysics, Julius Maximilian University Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Burgert
authors.affiliation: Department of Biotechnology and Biophysics, Julius Maximilian University Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Schlegel
authors.affiliation: Institute of Hygiene and Microbiology, Julius Maximilian University Würzburg, 97080, Würzburg, Germany.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Bécam
authors.affiliation: Department of Biotechnology and Biophysics, Julius Maximilian University Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Sören
authors.initials: S
authors.lastname: Doose
authors.affiliation: Department of Neuroscience and Regenerative Medicine, Medical College of Georgia, Augusta University, Augusta, GA, 30912, USA.
authors.forename: Erhard
authors.initials: E
authors.lastname: Bieberich
authors.affiliation: Institute of Hygiene and Microbiology, Julius Maximilian University Würzburg, 97080, Würzburg, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Schubert-Unkmeir
authors.affiliation: Department of Biotechnology and Biophysics, Julius Maximilian University Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Sauer
mesh_terms: D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002518:Ceramides / Q000378:metabolism*; D006801:Humans; D013269:Stochastic Processes
chemical_list: D002518:Ceramides
keywords: ceramide-rich platforms; ceramides; dSTORM; sphingolipids; super-resolution microscopy