Endocytosis of NBD‐Sphingolipids in Neurons: Exclusion from Degradative Compartments and Transport to the Golgi Complex
Sphingolipids are abundant constituents of neuronal membranes that have been implicated in intracellular signaling, neurite outgrowth and differentiation. Differential localization and trafficking of lipids to membrane domains contribute to the specialized functions. In non‐neuronal cultured cell li...
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Published in | Traffic (Copenhagen, Denmark) Vol. 2; no. 6; pp. 395 - 405 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
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Oxford, UK
Munksgaard International Publishers
01.06.2001
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Abstract | Sphingolipids are abundant constituents of neuronal membranes that have been implicated in intracellular signaling, neurite outgrowth and differentiation. Differential localization and trafficking of lipids to membrane domains contribute to the specialized functions. In non‐neuronal cultured cell lines, plasma membrane short‐chain sphingomyelin and glucosylceramide are recycled via endosomes or sorted to degradative compartments. However, depending on cell type and lipid membrane composition, short‐chain glucosylceramide can also be diverted to the Golgi complex. Here, we show that NBD‐labeled glucosylceramide and sphingomyelin are transported from the plasma membrane to the Golgi complex in cultured rat hippocampal neurons irrespective of the stage of neuronal differentiation. Golgi complex localization was confirmed by colocalization and Golgi disruption studies, and importantly did not result from conversion of NBD‐glucosylceramide or NBD‐sphingomyelin to NBD‐ceramide. Double‐labeling experiments with transferrin or wheat‐germ agglutinin showed that NBD‐sphingolipids are first internalized to early/recycling endosomes, and subsequently transported to the Golgi complex. The internalization of these two sphingolipid analogs was energy and temperature dependent, and their intracellular transport was insensitive to the NBD fluorescence quencher sodium dithionite. These results indicate that vesicles mediate the transport of internalized NBD‐glucosylceramide and NBD‐sphingomyelin to the Golgi complex. |
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AbstractList | Sphingolipids are abundant constituents of neuronal membranes that have been implicated in intracellular signaling, neurite outgrowth and differentiation. Differential localization and trafficking of lipids to membrane domains contribute to the specialized functions. In non‐neuronal cultured cell lines, plasma membrane short‐chain sphingomyelin and glucosylceramide are recycled via endosomes or sorted to degradative compartments. However, depending on cell type and lipid membrane composition, short‐chain glucosylceramide can also be diverted to the Golgi complex. Here, we show that NBD‐labeled glucosylceramide and sphingomyelin are transported from the plasma membrane to the Golgi complex in cultured rat hippocampal neurons irrespective of the stage of neuronal differentiation. Golgi complex localization was confirmed by colocalization and Golgi disruption studies, and importantly did not result from conversion of NBD‐glucosylceramide or NBD‐sphingomyelin to NBD‐ceramide. Double‐labeling experiments with transferrin or wheat‐germ agglutinin showed that NBD‐sphingolipids are first internalized to early/recycling endosomes, and subsequently transported to the Golgi complex. The internalization of these two sphingolipid analogs was energy and temperature dependent, and their intracellular transport was insensitive to the NBD fluorescence quencher sodium dithionite. These results indicate that vesicles mediate the transport of internalized NBD‐glucosylceramide and NBD‐sphingomyelin to the Golgi complex. |
Author | Kok, Jan Willem Saffrich, Rainer Babià, Teresa Egea, Gustavo Dotti, Carlos G. Ledesma, Maria Dolores |
Author_xml | – sequence: 1 givenname: Teresa surname: Babià fullname: Babià, Teresa – sequence: 2 givenname: Maria Dolores surname: Ledesma fullname: Ledesma, Maria Dolores – sequence: 3 givenname: Rainer surname: Saffrich fullname: Saffrich, Rainer – sequence: 4 givenname: Jan Willem surname: Kok fullname: Kok, Jan Willem – sequence: 5 givenname: Carlos G. surname: Dotti fullname: Dotti, Carlos G. – sequence: 6 givenname: Gustavo surname: Egea fullname: Egea, Gustavo |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/11389767$$D View this record in MEDLINE/PubMed |
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SubjectTerms | Animals Antineoplastic Agents - pharmacology Brefeldin A - pharmacology Cell Differentiation Cell Line Cell Membrane - metabolism Cells, Cultured Chromatography, Thin Layer Cytoplasm - metabolism Endocytosis Endosomes - metabolism Glucosylceramides - biosynthesis Golgi Apparatus - metabolism Golgi complex Hippocampus - cytology Hippocampus - metabolism lipid trafficking Lysosomes - metabolism Microscopy, Phase-Contrast NBD‐sphingolipids neurons Neurons - metabolism Nocodazole - pharmacology Protein Synthesis Inhibitors - pharmacology Rats Sphingolipids - biosynthesis Sphingomyelins - metabolism Temperature Time Factors Transferrin - metabolism |
Title | Endocytosis of NBD‐Sphingolipids in Neurons: Exclusion from Degradative Compartments and Transport to the Golgi Complex |
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