Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1982 Aug 1;94(2):279–286. doi: 10.1083/jcb.94.2.279

Pinocytosis in mouse L-fibroblasts: ultrastructural evidence for a direct membrane shuttle between the plasma membrane and the lysosomal compartment

PMCID: PMC2112883  PMID: 7107699

Abstract

Mouse L-fibroblasts internalized large amounts of cationized ferritin (CF) by pinocytosis. Initially (60-90 s after addition of CF to cell monolayers at 37 degrees C), CF was found in vesicles measuring 100-400 nm (sectioned diameter) and as small clusters adhering to the inner aspect of the limiting membrane of a few large (greater than 600 nm) vacuoles. After 5-30 min, CF labeling of large vacuoles was pronounced and continuous. Moreover, 70-80% of all labeled structures were tiny (less than 100 nm) vesicles. However, the absolute frequency of tiny vesicles increased more than twofold from 5 min to 30 min. When the cells were incubated with CF for 30 min, then washed and further incubated for 3 h without CF, almost all CF was present in dense bodies (100-500 nm). When L-cells were first incubated with horseradish peroxidase (HRP), then washed and incubated with CF, double-labeled vacuoles were observed. Tiny vesicles also contained HRP-CF, and small HRP-CF patches were localized on the cell surface. Distinct labeling of stacked Golgi cisterns was not observed in any experiment. These observations suggest that the numerous tiny vesicles are not endocytic but rather pinch off from the large vacuoles and move towards the cell surface to fuse with the plasma membrane. Thus, ultrastructural evidence is provided in favor of a direct membrane shuttle between the plasma membrane and the lysosomal compartment.

Full Text

The Full Text of this article is available as a PDF (3.3 MB).

Selected References

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

  1. Besterman J. M., Airhart J. A., Woodworth R. C., Low R. B. Exocytosis of pinocytosed fluid in cultured cells: kinetic evidence for rapid turnover and compartmentation. J Cell Biol. 1981 Dec;91(3 Pt 1):716–727. doi: 10.1083/jcb.91.3.716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borysenko J. Z., Woods W. Density, distribution and mobility of surface anions on a normal/transformed cell pair. Exp Cell Res. 1979 Feb;118(2):215–227. doi: 10.1016/0014-4827(79)90146-0. [DOI] [PubMed] [Google Scholar]
  3. Bowers B., Olszewski T. E. Pinocytosis in Acanthamoeba castellanii. Kinetics and morphology. J Cell Biol. 1972 Jun;53(3):681–694. doi: 10.1083/jcb.53.3.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Christensen E. I., Carone F. A., Rennke H. G. Effect of molecular charge on endocytic uptake of ferritin in renal proximal tubule cells. Lab Invest. 1981 Apr;44(4):351–358. [PubMed] [Google Scholar]
  5. Denef J. F., Ekholm R. Membrane labeling with cationized ferritin in isolated thyroid follicles. J Ultrastruct Res. 1980 May;71(2):203–221. doi: 10.1016/s0022-5320(80)90107-0. [DOI] [PubMed] [Google Scholar]
  6. Farquhar M. G., Palade G. E. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage. J Cell Biol. 1981 Dec;91(3 Pt 2):77s–103s. doi: 10.1083/jcb.91.3.77s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Grinnell F., Tobleman M. Q., Hackenbrock C. R. The distribution and mobility of anionic sites on the surfaces of baby hamster kidney cells. J Cell Biol. 1975 Sep;66(3):470–479. doi: 10.1083/jcb.66.3.470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Muller W. A., Steinman R. M., Cohn Z. A. The membrane proteins of the vacuolar system. II. Bidirectional flow between secondary lysosomes and plasma membrane. J Cell Biol. 1980 Jul;86(1):304–314. doi: 10.1083/jcb.86.1.304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Schneider Y. J., de Duve C., Trouet A. Fate of plasma membrane during endocytosis. III. Evidence for incomplete breakdown of immunoglobulin in lysosomes of cultured fibroblasts. J Cell Biol. 1981 Feb;88(2):380–387. doi: 10.1083/jcb.88.2.380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Skutelsky E., Hardy B. Regeneration of plasmalemma and surface properties in macrophages. Exp Cell Res. 1976 Sep;101(2):337–345. doi: 10.1016/0014-4827(76)90386-4. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Steinman R. M., Silver J. M., Cohn Z. A. Pinocytosis in fibroblasts. Quantitative studies in vitro. J Cell Biol. 1974 Dec;63(3):949–969. doi: 10.1083/jcb.63.3.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Takata K., Nishiyama F., Hirano H. Double labeling study of anionic sites and concanavalin A binding sites in monkey macrophages. J Histochem Cytochem. 1981 Jul;29(7):858–863. doi: 10.1177/29.7.6790605. [DOI] [PubMed] [Google Scholar]
  19. Tartakoff A., Vassalli P., Montesano R. Plasma cell endocytosis: is it related to immunoglobulin secretion? Eur J Cell Biol. 1981 Dec;26(1):188–197. [PubMed] [Google Scholar]
  20. Thilo L., Vogel G. Kinetics of membrane internalization and recycling during pinocytosis in Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1980 Feb;77(2):1015–1019. doi: 10.1073/pnas.77.2.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thyberg J. Internalization of cationized ferritin into the Golgi complex of cultured mouse peritoneal macrophages. Effects of colchicine and cytochalasin B. Eur J Cell Biol. 1980 Dec;23(1):95–103. [PubMed] [Google Scholar]
  22. Thyberg J., Nilsson J., Hellgren D. Recirculation of cationized ferritin in cultured mouse peritoneal macrophages. Electron microscopic and cytochemical studies with double-labeling technique. Eur J Cell Biol. 1980 Dec;23(1):85–94. [PubMed] [Google Scholar]
  23. Van Deurs B., Von Bülow F., Møller M. Vesicular transport of cationized ferritin by the epithelium of the rat choroid plexus. J Cell Biol. 1981 Apr;89(1):131–139. doi: 10.1083/jcb.89.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES