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• W2090551118 abstract "The intracellular flow and fate of two fluorescently labeled sphingolipids, 6-[N-(7-nitro-2,1,3benzoxadiazol-4-yl)amino]hexanoyl glucosyl sphingosine (C6-NBD-glucosylceramide) and C6-NBDsphingomyelin, was examined in the human colon adenocarcinoma cell line HT29. After their insertion into the plasma membrane at low temperature and subsequent warming of the cells to 37°C, both sphingolipid analogues were internalized by endocytosis, but their intracellular site of destination differed . After 30 min of internalization, C6-NBD-glucosylceramide was localized in the Golgi apparatus, as demonstrated by colocalization with fluorescently labeled ceramide, a Golgi complex marker, and by showing that monensininduced disruption of the Golgi structure was paralleled by a similar perturbation of the fluorescence distribution . By contrast, C6-NBD-sphingomyelin does SORTING and recycling are well-known phenomena in intracellular protein trafficking, occurring during biosynthesis and endocytosis (1, 11, 13) . The importance of these kind of processes in the concomitant intracellular flow of lipids is now also gradually emerging . Several sphingolipids have been shown to reappear at the plasma membrane by a recycling mechanism, after exogenous membrane insertion and subsequent internalization via the endocytic pathway (5, 6) . In polarized cells, sorting of two newly synthesized sphingolipids, derived from a common precursor, has been shown to occur, as reflected by lipid specific, outbound trafficking to apical, and basolateral membrane domains (14) . However, it remains to be resolved whether lipids are also subject to sorting during inbound cellular trafficking, i .e., in endocytic events . During endocytosis, an extensive flow ofmembranes takes place . To monitor the trafficking pathways of lipids during this process, advantage can be taken of the intracellular processing of certain ligand-receptor complexes . This processing involves delivery to endosomal compartments, sorting, and either recycling to the plasma membrane or further movement down the endocytic pathway towards the lysosomal system (1, 12) . In the present study, it is shown that two fluorescently laTeresa Babiâs present address is Autonomous University of Barcelona, Department of Biochemistry, Bellaterra, Spain . © The Rockefeller University Press, 0021-9525/91/07/231/9 $2 .00 The Journal of Cell Biology, Volume 114, Number 2, July 1991231-239 not colocalize with the tagged ceramide. Rather, a colocalization with ricin, which is internalized by endocytosis and predominantly reaches the lysosomes, was observed, indicating that the site of delivery of this lipid is restricted to endosomal/lysosomal compartments . Also, in monensin-treated cells no change in the distribution of fluorescence was observed . Thus, these results demonstrate that (sphingo)lipid sorting can occur in the endocytic pathway. Interestingly, the observed sorting phenomenon was specific for glucosylceramide, when compared to other glycolipids, while only undifferentiated HT29 cells displayed the different routing of the two lipids . In differentiated HT29 cells the internalization pathway of sphingomyelin and glucosylceramide was indistinguishable from that of transferrin . beled sphingolipids, C6-NBD-glucosylceramide and C6NBD-sphingomyelin, are sorted during internalization along the endocytic pathway in HT29 cells . Using transferrin (Tf)l and ricin (in conjunction with their receptors) as intracellular traffic markers, evidence is presented indicating that C6-NBD-glucosylceramide is transported to the Golgi apparatus, whereas C6-NBD-sphingomyelin follows the endocytic pathway, down to the endosomal/lysosomal system . Furthermore, when the trafficking ofthese sphingolipids was compared in an undifferentiated and a differentiated HT29 cell type, the latter being derived by clonal selection, it could be shown that this sorting phenomenon only occurred in the undifferentiated cell type . This distinct difference between the two cell types may imply that lipid transport routes and sorting phenomena are related to differentiation of cells . Materials and Methods" @default.
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• W2090551118 date "1991-10-01" @default.
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• W2090551118 title "Sorting of sphingolipids in the endocytic pathway of HT29 cells" @default.
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• W2090551118 doi "https://doi.org/10.1016/0962-8924(91)90025-5" @default.
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