Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Oct 1;91(1):270–280. doi: 10.1083/jcb.91.1.270

Evidence for the sorting of endocytic vesicle contents during the receptor-mediated transport of IgG across the newborn rat intestine

PMCID: PMC2111928  PMID: 7298722

Abstract

Fc receptors on the luminal membranes of intestinal epithelial cells in the neonatal rat mediate the vesicular transfer of functionally intact IgG from the intestinal lumen to the circulation. In addition, there is a low level of nonselective protein uptake, but in this case transfer does not occur. To determine whether a specialized class of endocytic vesicles could account for the selective transfer of IgG, mixtures of IgG conjugated to ferritin (IgG-Ft) and unconjugated horseradish peroxidase (HRP) were injected together into the proximal intestine of 10-d-old rats, and the cellular distribution of these two different tracers was determined by electron microscopy. Virtually all apical endocytic vesicles contained both tracers, indicating simultaneous uptake of both proteins within the same vesicle. However, only IgG-Ft bound to the apical plasma membrane, appeared within coated vesicles at the lateral cell surface, and was released from cells. HRP did not bind to the luminal membrane and was not transferred across cells but was confined to apical lysosomes as identified by acid phosphatase and aryl sulfatase activities. To test the possibility that the binding of IgG to its receptor stimulated endocytosis, HRP was used as a fluid volume tracer, and the amount of HRP taken up by cells in the presence and absence of IgG was measured morphologically and biochemically. The results demonstrate that endocytosis in these cells is constitutive and occurs at the same level in the absence of IgG. The evidence presented indicates that the principal selective mechanism for IgG transfer is the binding of IgG to its receptor during endocytosis. Continued binding to vesicle membranes appears to be required for successful transfer because unbound proteins are removed from the transport pathway before exocytosis. These results favor the proposal that IgG is transferred across cells as an IgG-receptor complex.

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. Abrahamson D. R., Powers A., Rodewald R. Intestinal absorption of immune complexes by neonatal rats: a route of antigen transfer from mother to young. Science. 1979 Nov 2;206(4418):567–569. doi: 10.1126/science.493961. [DOI] [PubMed] [Google Scholar]
  2. Anderson R. G., Brown M. S., Goldstein J. L. Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts. Cell. 1977 Mar;10(3):351–364. doi: 10.1016/0092-8674(77)90022-8. [DOI] [PubMed] [Google Scholar]
  3. Bensadoun A., Weinstein D. Assay of proteins in the presence of interfering materials. Anal Biochem. 1976 Jan;70(1):241–250. doi: 10.1016/s0003-2697(76)80064-4. [DOI] [PubMed] [Google Scholar]
  4. Borthistle B. K., Kubo R. T., Brown W. R., Grey H. M. Studies on receptors for IgG on epithelial cells of the rat intestine. J Immunol. 1977 Aug;119(2):471–476. [PubMed] [Google Scholar]
  5. Brambell F. W. The transmission of immunity from mother to young and the catabolism of immunoglobulins. Lancet. 1966 Nov 19;2(7473):1087–1093. doi: 10.1016/s0140-6736(66)92190-8. [DOI] [PubMed] [Google Scholar]
  6. Brown M. S., Goldstein J. L. Receptor-mediated endocytosis: insights from the lipoprotein receptor system. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3330–3337. doi: 10.1073/pnas.76.7.3330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans E. M., Wrigglesworth J. M., Burdett K., Pover W. F. Studies on epithelial cells isolated from guinea pig small intestine. J Cell Biol. 1971 Nov;51(21):452–464. doi: 10.1083/jcb.51.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. FARQUHAR M. G., PALADE G. E. Glomerular permeability. II. Ferritin transfer across the glomerular capillary wall in nephrotic rats. J Exp Med. 1961 Nov 1;114:699–716. doi: 10.1084/jem.114.5.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farquhar M. G. Recovery of surface membrane in anterior pituitary cells. Variations in traffic detected with anionic and cationic ferritin. J Cell Biol. 1978 Jun;77(3):R35–R42. doi: 10.1083/jcb.77.3.r35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldstein J. L., Brown M. S. Binding and degradation of low density lipoproteins by cultured human fibroblasts. Comparison of cells from a normal subject and from a patient with homozygous familial hypercholesterolemia. J Biol Chem. 1974 Aug 25;249(16):5153–5162. [PubMed] [Google Scholar]
  11. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  12. Haigler H. T., McKanna J. A., Cohen S. Rapid stimulation of pinocytosis in human carcinoma cells A-431 by epidermal growth factor. J Cell Biol. 1979 Oct;83(1):82–90. doi: 10.1083/jcb.83.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Herzog V., Farquhar M. G. Luminal membrane retrieved after exocytosis reaches most golgi cisternae in secretory cells. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5073–5077. doi: 10.1073/pnas.74.11.5073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Herzog V., Miller F. Membrane retrieval in epithelial cells of isolated thyroid follicles. Eur J Cell Biol. 1979 Aug;19(3):203–215. [PubMed] [Google Scholar]
  15. Hopsu-Havu V. K., Arstila A. U., Helminen H. J., Kalimo H. O. Improvements in the method for the electron microscopic localization of arylsulphatase activity. Histochemie. 1967;8(1):54–64. doi: 10.1007/BF00279874. [DOI] [PubMed] [Google Scholar]
  16. Jones E. A., Waldmann T. A. The mechanism of intestinal uptake and transcellular transport of IgG in the neonatal rat. J Clin Invest. 1972 Nov;51(11):2916–2927. doi: 10.1172/JCI107116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Louvard D. Apical membrane aminopeptidase appears at site of cell-cell contact in cultured kidney epithelial cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4132–4136. doi: 10.1073/pnas.77.7.4132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Masur S. K., Holtzman E., Schwartz I. L., Walter R. Correlation between pinocytosis and hydroosmosis induced by neurohypophyseal hormones and mediated by adenosine 3',5'-cyclic monophosphate. J Cell Biol. 1971 Jun;49(3):582–594. doi: 10.1083/jcb.49.3.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morris B., Morris R. Quantitative assessment of the transmission of labelled protein by the proximal and distal regions of the small intestine of young rats. J Physiol. 1976 Mar;255(3):619–634. doi: 10.1113/jphysiol.1976.sp011299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Morris B., Morris R. The absorption of 125I-labelled immunoglobulin G by different regions of the gut in young rats. J Physiol. 1974 Sep;241(3):761–770. doi: 10.1113/jphysiol.1974.sp010683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moxon L. A., Wild A. E. Localisation of proteins in coated micropinocytotic vesicles during transport across rabbit yolk sac endoderm. Cell Tissue Res. 1976 Aug 20;171(2):175–193. doi: 10.1007/BF00219405. [DOI] [PubMed] [Google Scholar]
  23. Ottosen P. D., Courtoy P. J., Farquhar M. G. Pathways followed by membrane recovered from the surface of plasma cells and myeloma cells. J Exp Med. 1980 Jul 1;152(1):1–19. doi: 10.1084/jem.152.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rodewald R. Distribution of immunoglobulin G receptors in the small intestine of the young rat. J Cell Biol. 1980 Apr;85(1):18–32. doi: 10.1083/jcb.85.1.18. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rodewald R. Intestinal transport of antibodies in the newborn rat. J Cell Biol. 1973 Jul;58(1):189–211. doi: 10.1083/jcb.58.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rodewald R. Selective antibody transport in the proximal small intestine of the neonatal rat. J Cell Biol. 1970 Jun;45(3):635–640. doi: 10.1083/jcb.45.3.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rodewald R. pH-dependent binding of immunoglobulins to intestinal cells of the neonatal rat. J Cell Biol. 1976 Nov;71(2):666–669. doi: 10.1083/jcb.71.2.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. SRI RAM J., TAWDE S. S., PIERCE G. B., Jr, MIDGLEY A. R., Jr Preparation of antibody-ferritin conjugates for immunoelectron microscopy. J Cell Biol. 1963 Jun;17:673–675. doi: 10.1083/jcb.17.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schneider Y. J., Tulkens P., de Duve C., Trouet A. Fate of plasma membrane during endocytosis. II. Evidence for recycling (shuttle) of plasma membrane constituents. J Cell Biol. 1979 Aug;82(2):466–474. doi: 10.1083/jcb.82.2.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Silverstein S. C., Steinman R. M., Cohn Z. A. Endocytosis. Annu Rev Biochem. 1977;46:669–722. doi: 10.1146/annurev.bi.46.070177.003321. [DOI] [PubMed] [Google Scholar]
  32. Steinman R. M., Brodie S. E., Cohn Z. A. Membrane flow during pinocytosis. A stereologic analysis. J Cell Biol. 1976 Mar;68(3):665–687. doi: 10.1083/jcb.68.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Steinman R. M., Cohn Z. A. The interaction of particulate horseradish peroxidase (HRP)-anti HRP immune complexes with mouse peritoneal macrophages in vitro. J Cell Biol. 1972 Dec;55(3):616–634. doi: 10.1083/jcb.55.3.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Steinman R. M., Cohn Z. A. The interaction of soluble horseradish peroxidase with mouse peritoneal macrophages in vitro. J Cell Biol. 1972 Oct;55(1):186–204. doi: 10.1083/jcb.55.1.186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wallace K. H., Rees A. R. Studies on the immunoglobulin-G Fc-fragment receptor from neonatal rat small intestine. Biochem J. 1980 Apr 15;188(1):9–16. doi: 10.1042/bj1880009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wild A. E., Richardson L. J. Direct evidence for pH-dependent Fc receptors on proximal enterocytes of suckling rat gut. Experientia. 1979 Jun 15;35(6):838–840. doi: 10.1007/BF01968284. [DOI] [PubMed] [Google Scholar]
  37. Willingham M. C., Pastan I. The receptosome: an intermediate organelle of receptor mediated endocytosis in cultured fibroblasts. Cell. 1980 Aug;21(1):67–77. doi: 10.1016/0092-8674(80)90115-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES