Use of BODIPY-labeled sphingolipids to study membrane traffic along the endocytic pathway.
In this chapter we discuss the use of BODIPY-labeled sphingolipids to study lipid transport along the endocytic pathway of cultured mammalian cells. The unique spectral properties of the BODIPY fluorophore allow the investigator to distinguish various populations of labeled endosome 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 particularly useful for visualizing the accumulation of lipids in the lysosomes of storage disease fibroblasts and may provide a useful method for screening various agents that abrogate this accumulation.
Department of Biochemistry and Molecular Biology, Mayo Clinic and Foundation, Rochester, Minnesota 55905, USA. pagano.richard@mayo.edu
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D000818:Animals; D001692:Biological Transport; D001896:Boron Compounds; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D004705:Endocytosis; D005456:Fluorescent Dyes; D006801:Humans; D008322:Mammals; D013106:Sphingolipidoses / Q000378:metabolism*; D013107:Sphingolipids / Q000378:metabolism*
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D013107:Sphingolipids