Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1985 Dec 1;101(6):2113–2123. doi: 10.1083/jcb.101.6.2113

Transcellular transport of polymeric IgA in the rat hepatocyte: biochemical and morphological characterization of the transport pathway

PMCID: PMC2113994  PMID: 4066752

Abstract

Polymeric IgA (pIgA) is transported by liver parenchymal cells (hepatocytes) from blood to bile via a receptor-mediated process. We have studied the intracellular pathway taken by a TEPC15 mouse myeloma pIgA. When from 1 microgram to 1 mg 125I-pIgA was injected into the saphenous vein of a rat, 36% was transported as intact protein into the bile over a 3-h period. The concentration of transported 125I-pIgA was maximal in bile 30-60 min after injection, and approximately 80% of the total 125I-pIgA ultimately transported had been secreted into bile by 90 min. A horseradish peroxidase-pIgA conjugate (125I-pIgA-HRP) was transported to a similar extent and with kinetics similar to that of unconjugated 125I-pIgA and was therefore used to visualize the transport pathway. Peroxidase cytochemistry of livers fixed in situ 2.5 to 10 min after 125I-pIgA-HRP injection demonstrated a progressive redistribution of labeled structures from the sinusoidal area to intermediate and bile canalicular regions of the hepatocyte cytoplasm. Although conjugate-containing structures began accumulating in the bile canalicular region at these early times, no conjugate was present in bile until 20 min. From 7.5 to 45 min after injection approximately 30% of the labeled structures were in regions that contained Golgi complexes and lysosomes; however, we found no evidence that either organelle contained 125I-pIgA-HRP. At least 85% of all positive structures in the hepatocyte were vesicles of 110-160-nm median diameters, with the remaining structures accounted for by tubules and multivesicular bodies. Vesicles in the bile canalicular region tended to be larger than those in the sinusoidal region. Serial sectioning showed that the 125I-pIgA-HRP-containing structures were relatively simple (predominantly vesicular) and that extensive interconnections did not exist between structures in the sinusoidal and bile canalicular regions.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bartles J. R., Braiterman L. T., Hubbard A. L. Endogenous and exogenous domain markers of the rat hepatocyte plasma membrane. J Cell Biol. 1985 Apr;100(4):1126–1138. doi: 10.1083/jcb.100.4.1126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandtzaeg P., Prydz H. Direct evidence for an integrated function of J chain and secretory component in epithelial transport of immunoglobulins. Nature. 1984 Sep 6;311(5981):71–73. doi: 10.1038/311071a0. [DOI] [PubMed] [Google Scholar]
  3. Brown W. J., Farquhar M. G. The mannose-6-phosphate receptor for lysosomal enzymes is concentrated in cis Golgi cisternae. Cell. 1984 Feb;36(2):295–307. doi: 10.1016/0092-8674(84)90223-x. [DOI] [PubMed] [Google Scholar]
  4. Dunn W. A., Hubbard A. L. Receptor-mediated endocytosis of epidermal growth factor by hepatocytes in the perfused rat liver: ligand and receptor dynamics. J Cell Biol. 1984 Jun;98(6):2148–2159. doi: 10.1083/jcb.98.6.2148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fisher M. M., Nagy B., Bazin H., Underdown B. J. Biliary transport of IgA: role of secretory component. Proc Natl Acad Sci U S A. 1979 Apr;76(4):2008–2012. doi: 10.1073/pnas.76.4.2008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Geuze H. J., Slot J. W., Strous G. J., Lodish H. F., Schwartz A. L. Intracellular site of asialoglycoprotein receptor-ligand uncoupling: double-label immunoelectron microscopy during receptor-mediated endocytosis. Cell. 1983 Jan;32(1):277–287. doi: 10.1016/0092-8674(83)90518-4. [DOI] [PubMed] [Google Scholar]
  7. Geuze H. J., Slot J. W., Strous G. J., Peppard J., von Figura K., Hasilik A., Schwartz A. L. Intracellular receptor sorting during endocytosis: comparative immunoelectron microscopy of multiple receptors in rat liver. Cell. 1984 May;37(1):195–204. doi: 10.1016/0092-8674(84)90315-5. [DOI] [PubMed] [Google Scholar]
  8. Halpern M. S., Coffman R. L. Polymer formation and J chain synthesis in mouse plasmacytomas. J Immunol. 1972 Oct;109(4):674–680. [PubMed] [Google Scholar]
  9. Hubbard A. L., Bartles J. R., Braiterman L. T. Identification of rat hepatocyte plasma membrane proteins using monoclonal antibodies. J Cell Biol. 1985 Apr;100(4):1115–1125. doi: 10.1083/jcb.100.4.1115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hubbard A. L., Ma A. Isolation of rat hepatocyte plasma membranes. II. Identification of membrane-associated cytoskeletal proteins. J Cell Biol. 1983 Jan;96(1):230–239. doi: 10.1083/jcb.96.1.230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hubbard A. L., Wilson G., Ashwell G., Stukenbrok H. An electron microscope autoradiographic study of the carbohydrate recognition systems in rat liver. I. Distribution of 125I-ligands among the liver cell types. J Cell Biol. 1979 Oct;83(1):47–64. doi: 10.1083/jcb.83.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kleinman R. E., Harmatz P. R., Walker W. A. The liver: an integral part of the enteric mucosal immune system. Hepatology. 1982 May-Jun;2(3):379–384. doi: 10.1002/hep.1840020315. [DOI] [PubMed] [Google Scholar]
  13. Lemaître-Coelho I., Altamirano G. A., Barranco-Acosta C., Meykens R., Vaerman J. P. In vivo experiments involving secretory component in the rat hepatic transfer of polymeric IgA from blood into bile. Immunology. 1981 Jun;43(2):261–270. [PMC free article] [PubMed] [Google Scholar]
  14. Lemaître-Coelho I., Jackson G. D., Vaerman J. P. Rat bile as a convenient source of secretory IgA and free secretory component. Eur J Immunol. 1977 Aug;7(8):588–590. doi: 10.1002/eji.1830070818. [DOI] [PubMed] [Google Scholar]
  15. Limet J. N., Schneider Y. J., Vaerman J. P., Trouet A. Interaction of rat IgA with cultured rat hepatocytes: binding site, drug effects. Toxicology. 1980;18(3):187–194. doi: 10.1016/0300-483x(80)90063-3. [DOI] [PubMed] [Google Scholar]
  16. Mostov K. E., Blobel G. A transmembrane precursor of secretory component. The receptor for transcellular transport of polymeric immunoglobulins. J Biol Chem. 1982 Oct 10;257(19):11816–11821. [PubMed] [Google Scholar]
  17. Mullock B. M., Luzio J. P., Hinton R. H. Preparation of a low-density species of endocytic vesicle containing immunoglobulin A. Biochem J. 1983 Sep 15;214(3):823–827. doi: 10.1042/bj2140823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Orlans E., Peppard J. V., Payne A. W., Fitzharris B. M., Mullock B. M., Hinton R. H., Hall J. G. Comparative aspects of the hepatobiliary transport of IgA. Ann N Y Acad Sci. 1983 Jun 30;409:411–427. doi: 10.1111/j.1749-6632.1983.tb26886.x. [DOI] [PubMed] [Google Scholar]
  19. Orlans E., Peppard J., Fry J. F., Hinton R. H., Mullock B. M. Secretory component as the receptor for polymeric IgA on rat hepatocytes. J Exp Med. 1979 Dec 1;150(6):1577–1581. doi: 10.1084/jem.150.6.1577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Orlans E., Peppard J., Reynolds J., Hall J. Rapid active transport of immunoglobulin A from blood to bile. J Exp Med. 1978 Feb 1;147(2):588–592. doi: 10.1084/jem.147.2.588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Peppard J., Orlans E., Payne A. W., Andrew E. The elimination of circulating complexes containing polymeric IgA by excretion in the bile. Immunology. 1981 Jan;42(1):83–89. [PMC free article] [PubMed] [Google Scholar]
  22. Potter M. Immunoglobulin-producing tumors and myeloma proteins of mice. Physiol Rev. 1972 Jul;52(3):631–719. doi: 10.1152/physrev.1972.52.3.631. [DOI] [PubMed] [Google Scholar]
  23. Renston R. H., Jones A. L., Christiansen W. D., Hradek G. T., Underdown B. J. Evidence for a vesicular transport mechanism in hepatocytes for biliary secretion of immunoglobulin A. Science. 1980 Jun 13;208(4449):1276–1278. doi: 10.1126/science.7375938. [DOI] [PubMed] [Google Scholar]
  24. Renston R. H., Maloney D. G., Jones A. L., Hradek G. T., Wong K. Y., Goldfine I. D. Bile secretory apparatus: evidence for a vesicular transport mechanism for proteins in the rat, using horseradish peroxidase and [125I]insulin. Gastroenterology. 1980 Jun;78(6):1373–1388. [PubMed] [Google Scholar]
  25. Roman L. M., Hubbard A. L. A domain-specific marker for the hepatocyte plasma membrane: localization of leucine aminopeptidase to the bile canalicular domain. J Cell Biol. 1983 Jun;96(6):1548–1558. doi: 10.1083/jcb.96.6.1548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schiff J. M., Fisher M. M., Underdown B. J. Receptor-mediated biliary transport of immunoglobulin A and asialoglycoprotein: sorting and missorting of ligands revealed by two radiolabeling methods. J Cell Biol. 1984 Jan;98(1):79–89. doi: 10.1083/jcb.98.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Socken D. J., Jeejeebhoy K. N., Bazin H., Underdown B. J. Identification of secretory component as an IgA receptor on rat hepatocytes. J Exp Med. 1979 Dec 1;150(6):1538–1548. doi: 10.1084/jem.150.6.1538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Steer C. J., Ashwell G. Studies on a mammalian hepatic binding protein specific for asialoglycoproteins. Evidence for receptor recycling in isolated rat hepatocytes. J Biol Chem. 1980 Apr 10;255(7):3008–3013. [PubMed] [Google Scholar]
  29. Sztul E. S., Howell K. E., Palade G. E. Biogenesis of the polymeric IgA receptor in rat hepatocytes. I. Kinetic studies of its intracellular forms. J Cell Biol. 1985 Apr;100(4):1248–1254. doi: 10.1083/jcb.100.4.1248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sztul E. S., Howell K. E., Palade G. E. Biogenesis of the polymeric IgA receptor in rat hepatocytes. II. Localization of its intracellular forms by cell fractionation studies. J Cell Biol. 1985 Apr;100(4):1255–1261. doi: 10.1083/jcb.100.4.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sztul E. S., Howell K. E., Palade G. E. Intracellular and transcellular transport of secretory component and albumin in rat hepatocytes. J Cell Biol. 1983 Nov;97(5 Pt 1):1582–1591. doi: 10.1083/jcb.97.5.1582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Takahashi I., Nakane P. K., Brown W. R. Ultrastructural events in the translocation of polymeric IgA by rat hepatocytes. J Immunol. 1982 Mar;128(3):1181–1187. [PubMed] [Google Scholar]
  33. Tanabe T., Pricer W. E., Jr, Ashwell G. Subcellular membrane topology and turnover of a rat hepatic binding protein specific for asialoglycoproteins. J Biol Chem. 1979 Feb 25;254(4):1038–1043. [PubMed] [Google Scholar]
  34. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wall D. A., Hubbard A. L. Galactose-specific recognition system of mammalian liver: receptor distribution on the hepatocyte cell surface. J Cell Biol. 1981 Sep;90(3):687–696. doi: 10.1083/jcb.90.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wall D. A., Wilson G., Hubbard A. L. The galactose-specific recognition system of mammalian liver: the route of ligand internalization in rat hepatocytes. Cell. 1980 Aug;21(1):79–93. doi: 10.1016/0092-8674(80)90116-6. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES