Use of N-[5-(5,7-dimethyl boron dipyrromethene difluoride-sphingomyelin to study membrane traffic along the endocytic pathway.
We have used N-[5-(5,7-dimethyl boron dipyrromethene difluoride)-1-pentanoyl]-D-erythro-sphingosylphosphorylcholine (C5-DMB-SM or 'BODIPY-SM'), a fluorescent analog of sphingomyelin, to study lipid transport along the endocytic pathway of human skin fibroblasts. The unique spectral properties of the BODIPY fluorophore allow the investigator to distinguish various populations of labeled endosomes and lysosomes within the living cell by fluorescence microscopy, and in conjunction with quantitative fluorescence microscopy, to estimate the concentration of these lipids in different intracellular compartments. This methodology is also applicable for visualizing the accumulation of lipids in the endosomes and lysosomes of storage disease fibroblasts.
Pagano
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RE
Department of Biochemistry and Molecular Biology, Thoracic Diseases Research Unit, Mayo Clinic and Foundation, Rochester, MN 55905-0001, USA. pagano.richard@mayo.edu
Watanabe
R
R
Wheatley
C
C
Chen
C S
CS
D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D011992:Endosomes / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000493:pharmacokinetics*; D006801:Humans; D050356:Lipid Metabolism; D008247:Lysosomes / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009542:Niemann-Pick Diseases / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D013109:Sphingomyelins / Q000493:pharmacokinetics*
D005456:Fluorescent Dyes; C105705:N-(5-(5,7-dimethyl Bodipy)-1-pentanoyl)-erythro-sphingosylphosphorylcholine; D013109:Sphingomyelins