Journal of Cell Science, Vol 103, Issue 4 1139-1152, Copyright © 1992 by Company of Biologists

JOURNAL ARTICLES

Recycling pathways of glucosylceramide in BHK cells: distinct involvement of early and late endosomes

JW Kok, K Hoekstra, S Eskelinen and D Hoekstra
University of Groningen, Laboratory of Physiological Chemistry, The Netherlands.

Recycling pathways of the sphingolipid glucosylceramide were studied by employing a fluorescent analog of glucosylceramide, 6(-)[N-(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]hexanoylglucosyl sphingosine (C6-NBD-glucosylceramide). Direct recycling of the glycolipid from early endosomes to the plasma membrane occurs, as could be shown after treating the cells with the microtubule-disrupting agent nocodazole, which causes inhibition of the glycolipid's trafficking from peripheral early endosomes to centrally located late endosomes. When the microtubuli are intact, at least part of the glucosylceramide is transported from early to late endosomes together with ricin. Interestingly, also N-(lissamine rhodamine B sulfonyl)phosphatidylethanolamine (N-Rh-PE), a membrane marker of the fluid-phase endocytic pathway, is transported to this endosomal compartment. However, in contrast to both ricin and N-Rh-PE, the glucosylceramide can escape from this organelle and recycle to the plasma membrane. Monensin and brefeldin A have little effect on this recycling pathway, which would exclude extensive involvement of early Golgi compartments in recycling. Hence, the small fraction of the glycolipid that colocalizes with transferrin (Tf) in the Golgi area might directly recycle via the trans-Golgi network. When the intracellular pH was lowered to 5.5, recycling was drastically reduced, in accordance with the impeding effect of low intracellular pH on vesicular transport during endocytosis and in the biosynthetic pathway. Our results thus demonstrate the existence of at least two recycling pathways for glucosylceramide and indicate the relevance of early endosomes in recycling of both proteins and lipids.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				Y. Sagiv, K. Hudspeth, J. Mattner, N. Schrantz, R. K. Stern, D. Zhou, P. B. Savage, L. Teyton, and A. Bendelac Cutting Edge: Impaired Glycosphingolipid Trafficking and NKT Cell Development in Mice Lacking Niemann-Pick Type C1 Protein J. Immunol., July 1, 2006; 177(1): 26 - 30. [Abstract] [Full Text] [PDF]

				M. Husain and B. Moss Intracellular Trafficking of a Palmitoylated Membrane-Associated Protein Component of Enveloped Vaccinia Virus J. Virol., August 15, 2003; 77(16): 9008 - 9019. [Abstract] [Full Text] [PDF]

				V. Sriram, S. Cho, P. Li, P. W. O'Donnell, C. Dunn, K. Hayakawa, J. S. Blum, and R. R. Brutkiewicz Inhibition of glycolipid shedding rescues recognition of a CD1+ T cell lymphoma by natural killer T (NKT) cells PNAS, June 11, 2002; 99(12): 8197 - 8202. [Abstract] [Full Text] [PDF]

				F. Shi, A. Nomden, V. Oberle, Jan. B. F. N. Engberts, and D. Hoekstra Efficient cationic lipid-mediated delivery of antisense oligonucleotides into eukaryotic cells: down-regulation of the corticotropin-releasing factor receptor Nucleic Acids Res., May 15, 2001; 29(10): 2079 - 2087. [Abstract] [Full Text] [PDF]

				S. C.D. van IJzendoorn and D. Hoekstra (Glyco)sphingolipids Are Sorted in Sub-Apical Compartments in HepG2 Cells: A Role for Non-Golgi-Related Intracellular Sites in the Polarized Distribution of (Glyco)sphingolipids J. Cell Biol., August 10, 1998; 142(3): 683 - 696. [Abstract] [Full Text] [PDF]

				J. W. Kok, T. Babia, C. M. Filipeanu, A. Nelemans, G. Egea, and D. Hoekstra PDMP Blocks Brefeldin A-induced Retrograde Membrane Transport from Golgi to ER: Evidence for Involvement of Calcium Homeostasis and Dissociation from Sphingolipid Metabolism J. Cell Biol., July 13, 1998; 142(1): 25 - 38. [Abstract] [Full Text] [PDF]

				M. M. P. Zegers, K. J. M. Zaal, S. C. D. van IJzendoorn, K. Klappe, and D. Hoekstra Actin Filaments and Microtubules are Involved in Different Membrane Traffic Pathways That Transport Sphingolipids to the Apical Surface of Polarized HepG2 Cells Mol. Biol. Cell, July 1, 1998; 9(7): 1939 - 1949. [Abstract] [Full Text]

				J. W. Kok, M. Nikolova-Karakashian, K. Klappe, C. Alexander, and A. H. Merrill Jr. Dihydroceramide Biology. STRUCTURE-SPECIFIC METABOLISM AND INTRACELLULAR LOCALIZATION J. Biol. Chem., August 22, 1997; 272(34): 21128 - 21136. [Abstract] [Full Text] [PDF]

				R. T. Watson, S. Shigematsu, S.-H. Chiang, S. Mora, M. Kanzaki, I. G. Macara, A. R. Saltiel, and J. E. Pessin Lipid raft microdomain compartmentalization of TC10 is required for insulin signaling and GLUT4 translocation J. Cell Biol., August 20, 2001; 154(4): 829 - 840. [Abstract] [Full Text] [PDF]

				V. Puri, R. Watanabe, R. D. Singh, M. Dominguez, J. C. Brown, C. L. Wheatley, D. L. Marks, and R. E. Pagano Clathrin-dependent and -independent internalization of plasma membrane sphingolipids initiates two Golgi targeting pathways J. Cell Biol., August 6, 2001; 154(3): 535 - 548. [Abstract] [Full Text] [PDF]