TY - JOUR
T1 - Intracellular localization of organized lipid domains of C16-ceramide/cholesterol
AU - Goldschmidt-Arzi, Michal
AU - Shimoni, Eyal
AU - Sabanay, Helena
AU - Futerman, Anthony H.
AU - Addadi, Lia
N1 - Israel Science Foundation [1735/07]This work was supported by the Israel Science Foundation (1735/07). We thank Prof. Zvi Kam for help with image processing. Electron microscopy was performed at the Irving and Cherna Moskowitz Center for Nano and Bio-Nano Imaging at the Weizmann Institute. A.H. Futerman is the Joseph Meyerhoff Professor of Biochemistry and Lia Addadi is the incumbent of the Dorothy-and-Patrick-Gorman Professorial Chair of Biological Ultrastructure at the Weizmann Institute of Science.
PY - 2011/7
Y1 - 2011/7
N2 - Lipid microdomains, also called lipid rafts, consisting of sphingolipids and cholesterol, play important roles in membrane trafficking and in signaling. Despite years of study of the composition, size, half-life and dynamic organization of these domains, many open questions remain about their precise characteristics. To address some of these issues, we have developed a new experimental approach involving the use of specific monoclonal antibodies as recognition tools. One such antibody was raised against a homogeneous, mixed, ordered monolayer phase comprised of 60:40 mol% cholesterol:C16-ceramide, and has been used previously to demonstrate the existence of C16-ceramide/cholesterol domains in the membranes of cultured cells. We now use a combination of quantitative fluorescence microscopy, immuno-transmission electron microscopy and immuno-scanning cryo-electron microscopy, optimized for the study of intracellular lipid antigens. In a variety of cultured cells, C16-ceramide/cholesterol structural domains were found at high levels in late endosomes and in the trans-Golgi network, but were not found at statistically significant levels in early endosomes, lysosomes or the endoplasmic reticulum. We discuss the relevance of these results to understanding the role of lipid lateral organization in biological membranes. (C) 2011 Elsevier Inc. All rights reserved.
AB - Lipid microdomains, also called lipid rafts, consisting of sphingolipids and cholesterol, play important roles in membrane trafficking and in signaling. Despite years of study of the composition, size, half-life and dynamic organization of these domains, many open questions remain about their precise characteristics. To address some of these issues, we have developed a new experimental approach involving the use of specific monoclonal antibodies as recognition tools. One such antibody was raised against a homogeneous, mixed, ordered monolayer phase comprised of 60:40 mol% cholesterol:C16-ceramide, and has been used previously to demonstrate the existence of C16-ceramide/cholesterol domains in the membranes of cultured cells. We now use a combination of quantitative fluorescence microscopy, immuno-transmission electron microscopy and immuno-scanning cryo-electron microscopy, optimized for the study of intracellular lipid antigens. In a variety of cultured cells, C16-ceramide/cholesterol structural domains were found at high levels in late endosomes and in the trans-Golgi network, but were not found at statistically significant levels in early endosomes, lysosomes or the endoplasmic reticulum. We discuss the relevance of these results to understanding the role of lipid lateral organization in biological membranes. (C) 2011 Elsevier Inc. All rights reserved.
KW - Cell membrane
KW - Ceramide
KW - Electron microscopy
KW - Late endosomes
KW - Lipid domains
KW - Lipid rafts
UR - http://www.scopus.com/inward/record.url?scp=79956190674&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.jsb.2011.03.021
DO - https://doi.org/10.1016/j.jsb.2011.03.021
M3 - مقالة
C2 - 21473916
SN - 1047-8477
VL - 175
SP - 21
EP - 30
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 1
ER -