{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T06:31:17Z","timestamp":1779345077991,"version":"3.51.4"},"reference-count":52,"publisher":"Rockefeller University Press","issue":"3","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1998,8,10]]},"abstract":"<jats:p>In polarized HepG2 cells, the fluorescent sphingolipid analogues of glucosylceramide (C6-NBD-GlcCer) and sphingomyelin (C6-NBD-SM) display a preferential localization at the apical and basolateral domain, respectively, which is expressed during apical to basolateral transcytosis of the lipids (van IJzendoorn, S.C.D., M.M.P. Zegers, J.W. Kok, and D. Hoekstra. 1997. J. Cell Biol. 137:347\u2013457). In the present study we have identified a non-Golgi\u2013related, sub-apical compartment (SAC), in which sorting of the lipids occurs. Thus, in the apical to basolateral transcytotic pathway both C6-NBD-GlcCer and C6-NBD-SM accumulate in SAC at 18\u00b0C. At this temperature, transcytosing IgA also accumulates, and colocalizes with the lipids. Upon rewarming the cells to 37\u00b0C, the lipids are transported from the SAC to their preferred membrane domain. Kinetic evidence is presented that shows in a direct manner that after leaving SAC, sphingomyelin disappears from the apical region of the cell, whereas GlcCer is transferred to the apical, bile canalicular membrane. The sorting event is very specific, as the GlcCer epimer C6-NBD-galactosylceramide, like C6-NBD-SM, is sorted in the SAC and directed to the basolateral surface. It is demonstrated that transport of the lipids to and from SAC is accomplished by a vesicular mechanism, and is in part microtubule dependent. Furthermore, the SAC in HepG2 bear analogy to the apical recycling compartments, previously described in MDCK cells. However, in contrast to the latter, the structural integrity of SAC does not depend on an intact microtubule system. Taken together, we have identified a non-Golgi\u2013related compartment, acting as a \u201ctraffic center\u201d in apical to basolateral trafficking and vice versa, and directing the polarized distribution of sphingolipids in hepatic cells.<\/jats:p>","DOI":"10.1083\/jcb.142.3.683","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T16:46:23Z","timestamp":1027701983000},"page":"683-696","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":64,"title":["(Glyco)sphingolipids Are Sorted in Sub-Apical Compartments in HepG2 Cells: A Role for Non-Golgi\u2013Related Intracellular Sites in the Polarized Distribution of (Glyco)sphingolipids"],"prefix":"10.1083","volume":"142","author":[{"given":"Sven C.D.","family":"van IJzendoorn","sequence":"first","affiliation":[{"name":"Department of Physiological Chemistry, Faculty of Medical Sciences, University of Groningen, Groningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dick","family":"Hoekstra","sequence":"additional","affiliation":[{"name":"Department of Physiological Chemistry, Faculty of Medical Sciences, University of Groningen, Groningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[1998,8,10]]},"reference":[{"key":"2023072900041212900_B1","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1042\/bj2710193","article-title":"Priority targeting of gucosyl-phosphatidylinositol-anchored proteins to the bile canalicular (apical) plasma membrane of hepatocytes","volume":"271","author":"Ali","year":"1990","journal-title":"Biochem J"},{"key":"2023072900041212900_B2","doi-asserted-by":"crossref","first-page":"1877","DOI":"10.1172\/JCI115211","article-title":"The polymeric immunoglobulin receptor: a model protein to study transcytosis","volume":"87","author":"Apodaca","year":"1991","journal-title":"J Clin Invest"},{"key":"2023072900041212900_B3","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1083\/jcb.125.1.67","article-title":"Receptor mediated endocytosis of IgA in MDCK cells is via apical recycling endosomes","volume":"125","author":"Apodaca","year":"1994","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B4"},{"key":"2023072900041212900_B5","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1083\/jcb.124.1.83","article-title":"Basolateral to apical transcytosis in polarized cells is indirect and involves BFA and trimeric G protein sensitive passage through the apical endosome","volume":"124","author":"Barosso","year":"1994","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B6","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/0016-5085(93)90734-T","article-title":"Newly synthesized hepatocyte plasma membrane proteins are transported in transcytotic vesicles in the bile duct-ligated rat","volume":"105","author":"Barr","year":"1993","journal-title":"Gastroenterology"},{"key":"2023072900041212900_B7","doi-asserted-by":"crossref","first-page":"27834","DOI":"10.1074\/jbc.270.46.27834","article-title":"Immunoadsorption of hepatic vesicles carrying newly synthesized dipeptidyl peptidase IV and polymeric IgA receptor","volume":"270","author":"Barr","year":"1995","journal-title":"J Biol Chem"},{"key":"2023072900041212900_B8","doi-asserted-by":"crossref","first-page":"1241","DOI":"10.1083\/jcb.105.3.1241","article-title":"Biogenesis of the rat hepatocyte plasma membrane in vivo: Comparison of the pathways taken by apical and basolateral proteins using subcellular fractionation","volume":"105","author":"Bartles","year":"1987","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B9","first-page":"911","article-title":"A rapid method of total lipid extraction and purification","volume":"37","author":"Bligh","year":"1959","journal-title":"Can J Biochem Biophys"},{"key":"2023072900041212900_B10","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1083\/jcb.109.2.475","article-title":"Postendocytic sorting of the ligand for the polymeric immunoglobulin receptor in Madin-Darby canine kidney cells","volume":"109","author":"Breitfeld","year":"1989","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B11","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/0092-8674(92)90189-J","article-title":"Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical surface","volume":"68","author":"Brown","year":"1992","journal-title":"Cell"},{"key":"2023072900041212900_B12","first-page":"1021","article-title":"Plasma membrane domain localization and transcytosis of the glucagon-induced hepatic system A carrier","volume":"263","author":"Cariappa","year":"1992","journal-title":"Am J Physiol"},{"key":"2023072900041212900_B13","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/0092-8674(84)90315-5","article-title":"Intracellular receptor sorting during endocytosis: comparative immunoelectron microscopy of multiple receptors in rat liver","volume":"37","author":"Geuze","year":"1984","journal-title":"Cell"},{"key":"2023072900041212900_B14","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/0005-2760(87)90103-2","article-title":"Hydrolysis of galactosylsphingosine and lactosylsphingosine by \u03b2-galactosidases in human brain and cultured fibroblasts","volume":"920","author":"Goda","year":"1987","journal-title":"Biochim Biophys Acta"},{"key":"2023072900041212900_B15","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1016\/0955-0674(95)80013-1","article-title":"Membrane transport in the endocytic pathway","volume":"7","author":"Gruenberg","year":"1995","journal-title":"Curr Opin Cell Biol"},{"key":"2023072900041212900_B16","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1083\/jcb.108.4.1301","article-title":"Characterization of the early endosome and putative endocytic carrier vesicles in vivo and with an assay of vesicle fusion in vitro","volume":"108","author":"Gruenberg","year":"1989","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B17","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1242\/jcs.109.6.1215","article-title":"The transcytotic pathway of an apical plasma membrane protein (B10) in hepatocytes is similar to that of IgA and occurs via a tubular pericentriolar compartment","volume":"109","author":"Hemery","year":"1996","journal-title":"J Cell Sci"},{"key":"2023072900041212900_B18","doi-asserted-by":"crossref","first-page":"2113","DOI":"10.1083\/jcb.101.6.2113","article-title":"Transcellular transport of polymeric IgA in the rat hepatocyte: Biochemical and morphological characterization of the transport pathway","volume":"101","author":"Hoppe","year":"1985","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B19","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1083\/jcb.110.2.337","article-title":"Endocytic pathways in polarized Caco-2 cells: Identification of an endosomal compartment accessible from both apical and basolateral surfaces","volume":"110","author":"Hughson","year":"1990","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B20","doi-asserted-by":"crossref","first-page":"3291","DOI":"10.1083\/jcb.109.6.3291","article-title":"Expression of macrophage-lymphocyte Fc receptors in Madin-Darby canine kidney cells: Polarity and transcytosis differ for isoforms with or without coated pit localization domains","volume":"109","author":"Hunziker","year":"1989","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B21","doi-asserted-by":"crossref","first-page":"3515","DOI":"10.1002\/j.1460-2075.1990.tb07560.x","article-title":"Differential microtubule requirements for transcytosis in MDCK cells","volume":"9","author":"Hunziker","year":"1990","journal-title":"EMBO (Eur Mol Biol Organ) J"},{"key":"2023072900041212900_B22","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1083\/jcb.141.1.115","article-title":"Apical plasma membrane proteins and endolyn-78 travel through a sub-apical compartment in polarized WIF-B hepatocytes","volume":"141","author":"Ihrke","year":"1998","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B23","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1002\/hep.1840040613","article-title":"The fate of polymeric and secretory immunoglobulin A after retrograde infusion into the common bile duct in rats","volume":"4","author":"Jones AL, G.T. Hradek, D.L. Schmucker, and B.J. Underdown","year":"1984","journal-title":"Hepatology"},{"key":"2023072900041212900_B24","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/0009-3084(75)90029-8","article-title":"A facile synthesis of ceramides","volume":"15","author":"Kishimoto","year":"1975","journal-title":"Chem Phys Lipids"},{"key":"2023072900041212900_B25","doi-asserted-by":"crossref","first-page":"1139","DOI":"10.1242\/jcs.103.4.1139","article-title":"Recycling pathways of glucosylceramide in BHK cells: distinct involvement of early and late endosomes","volume":"103","author":"Kok","year":"1992","journal-title":"J Cell Sci"},{"key":"2023072900041212900_B26","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1083\/jcb.100.1.27","article-title":"Intracellular translocation of fluorescent sphingolipids in cultured fibroblasts: endogenously synthesized sphingomyelin and glucocerebroside analogues pass through the Golgi apparatus en route to the plasma membrane","volume":"100","author":"Lipsky","year":"1985","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B27","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1083\/jcb.121.3.553","article-title":"Rab17, a novel small GTPase, is specific for epithelial cells and is induced during cell polarization","volume":"121","author":"Lutcke","year":"1993","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B28","doi-asserted-by":"crossref","first-page":"2131","DOI":"10.1083\/jcb.99.6.2131","article-title":"Sorting of an apical plasma membrane glycoprotein occurs before it reaches the cell surface in cultured epithelial cells","volume":"99","author":"Matlin","year":"1984","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B29","doi-asserted-by":"crossref","first-page":"3163","DOI":"10.1002\/j.1460-2075.1990.tb07514.x","article-title":"Microtubule perturbation retards both the direct and the indirect apical pathway bus does not affect sorting of plasma membrane proteins in intestinal epithelial cells (Caco-2)","volume":"9","author":"Matter","year":"1990","journal-title":"EMBO (Eur Mol Biol Organ) J"},{"key":"2023072900041212900_B30","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.1083\/jcb.105.4.1595","article-title":"Receptor-mediated vectorial transcytosis of epidermal growth factor by Madin-Darby canine kidney cells","volume":"105","author":"Maratos-Flier","year":"1987","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B31","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1016\/0092-8674(84)90460-4","article-title":"Biogenesis of epithelial cell polarity: intracellular sorting and vectorial exocytosis of an apical plasma membrane glycoprotein","volume":"39","author":"Misek","year":"1984","journal-title":"Cell"},{"key":"2023072900041212900_B32","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1016\/0092-8674(86)90887-1","article-title":"Polymeric IgA receptor expressed in MDCK cells transcytoses IgA","volume":"46","author":"Mostov","year":"1986","journal-title":"Cell"},{"key":"2023072900041212900_B33","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1101\/SQB.1995.060.01.083","article-title":"Regulation of protein traffic in polarized epithelial cells: the polymeric immunoglobulin receptor model","volume":"60","author":"Mostov","year":"1995","journal-title":"Cold Spring Harbor Symp Quant Biol"},{"key":"2023072900041212900_B34","doi-asserted-by":"crossref","first-page":"3259","DOI":"10.1083\/jcb.109.6.3259","article-title":"Meeting of the apical and basolateral endocytic pathways of the Madin-Darby canine kidney cell in late endosomes","volume":"109","author":"Parton","year":"1989","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B35","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1083\/jcb.101.2.470","article-title":"Intracellular sorting and basolateral appearance of the G protein of vesicular stomatitis virus in Madin-Darby canine kidney cells","volume":"101","author":"Pfeiffer","year":"1985","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B36","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1053\/gast.1997.v112.pm9024305","article-title":"Increased levels of the multidrug resistance protein in lateral membranes of proliferating hepatocyte-derived cells","volume":"112","author":"Roelofsen","year":"1997","journal-title":"Gastroenterology"},{"key":"2023072900041212900_B37","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1083\/jcb.119.5.1173","article-title":"5\u2032 nucleotidase is sorted to the apical domain of hepatocytes via an indirect route","volume":"119","author":"Schell","year":"1992","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B38","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1146\/annurev.cb.01.110185.001331","article-title":"Cell surface polarity in epithelia","volume":"1","author":"Simons","year":"1985","journal-title":"Annu Rev Cell Biol"},{"key":"2023072900041212900_B39","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1038\/42408","article-title":"Functional rafts in cell membranes","volume":"387","author":"Simons","year":"1997","journal-title":"Nature"},{"key":"2023072900041212900_B40","doi-asserted-by":"crossref","first-page":"6197","DOI":"10.1021\/bi00417a001","article-title":"Lipid sorting in epithelial cells","volume":"27","author":"Simons","year":"1988","journal-title":"Biochemistry"},{"key":"2023072900041212900_B41","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/0092-8674(90)90357-K","article-title":"Polarized sorting in epithelia","volume":"62","author":"Simons","year":"1990","journal-title":"Cell"},{"key":"2023072900041212900_B42","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1016\/0016-5085(88)90252-1","article-title":"Receptor-mediated binding and uptake of immunoglobulin A by human liver","volume":"94","author":"Tomana","year":"1988","journal-title":"Gastroenterology"},{"key":"2023072900041212900_B43","doi-asserted-by":"crossref","first-page":"2081","DOI":"10.1083\/jcb.109.5.2081","article-title":"The response of the Golgi complex to microtubule alterations: The roles of metabolic energy and membrane traffic in Golgi complex organization","volume":"109","author":"Turner","year":"1989","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B44","doi-asserted-by":"crossref","first-page":"1160","DOI":"10.1073\/pnas.94.4.1160","article-title":"Novel pyridinium surfactants for efficient, nontoxic in vitro gene delivery","volume":"94","author":"van der Woude","year":"1997","journal-title":"Proc Natl Acad Sci USA"},{"key":"2023072900041212900_B45","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1083\/jcb.131.3.645","article-title":"Differential targeting of glucosylceramide and galactocylceramide analogues after synthesis but not during transcytosis in Madin-Darby canine kidney cells","volume":"131","author":"van Genderen","year":"1995","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B46","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/S0092-8674(00)81370-7","article-title":"MDR1 P-Glycoprotein is a lipid translocase of broad specificity, while MDR3 P-Glycoprotein specifically translocates phosphatidylcholine","volume":"87","author":"van Helvoort","year":"1996","journal-title":"Cell"},{"key":"2023072900041212900_B47","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1083\/jcb.137.2.347","article-title":"Segregation of glucosylceramide and sphingomyelin occurs in the apical to basolateral transcytotic route in HepG2 cells","volume":"137","author":"van IJzendoorn","year":"1997","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B48","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1083\/jcb.111.3.977","article-title":"Generation of lipid polarity in intestinal epithelial (Caco-2) cells: Sphingolipid synthesis in the Golgi complex and sorting before vesicular traffic to the plasma membrane","volume":"111","author":"van't Hof","year":"1990","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B49","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/S0962-8924(97)01130-6","article-title":"Apical targeting in polarized epithelial cells: there's more afloat than rafts","volume":"7","author":"Weimbs","year":"1997","journal-title":"Trends Cell Biol"},{"key":"2023072900041212900_B50","first-page":"10","article-title":"Intracellular sites involved in the biogenesis of bile canaliculi in hepatic cells","volume":"63","author":"Zaal","year":"1993","journal-title":"Eur J Cell Biol"},{"key":"2023072900041212900_B51","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1083\/jcb.140.5.1039","article-title":"Rab17 regulates membrane traffic through apical recycling endosomes in polarized epithelial cells","volume":"140","author":"Zacchi","year":"1998","journal-title":"J Cell Biol"},{"key":"2023072900041212900_B52","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1083\/jcb.138.2.307","article-title":"Sphingolipid transport to the apical plasma membrane in human hepatoma cells is controlled by PKC and PKA activity: A correlation with cell polarity in HepG2 cells","volume":"138","author":"Zegers","year":"1997","journal-title":"J Cell Biol"}],"container-title":["The Journal of Cell Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/142\/3\/683\/1846918\/9802111.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/142\/3\/683\/1846918\/9802111.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,29]],"date-time":"2023-07-29T00:05:32Z","timestamp":1690589132000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jcb\/article\/142\/3\/683\/15834\/Glyco-sphingolipids-Are-Sorted-in-Sub-Apical"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,8,10]]},"references-count":52,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1998,8,10]]}},"URL":"https:\/\/doi.org\/10.1083\/jcb.142.3.683","relation":{},"ISSN":["0021-9525","1540-8140"],"issn-type":[{"value":"0021-9525","type":"print"},{"value":"1540-8140","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,8,10]]}}}