Cell and Tissue Research

, Volume 222, Issue 3, pp 523–530 | Cite as

Distribution of phospholipids labeled with 3H-choline and relationship between membranous organelles in amoebae, as studied by electron-microscopic radioautography

  • Charles J. Flickinger
  • Graham A. Read
Article
Accepted:

Summary

Amoebae were injected with a solution of 3H-choline, and samples were prepared for electron-microscopic radioautography at intervals between 15 min and 24 h thereafter. At the earliest interval, the rough endoplasmic reticulum was the most heavily labeled organelle. At subsequent intervals, the proportion of silver grains over the rough endoplasmic reticulum decreased rapidly, while that associated with other membranes increased. Most notably this involved a rapid rise in labeling of vacuoles, up to 1 h, and a more gradual increase in plasma membrane labeling up to 2 h. The results suggest that phospholipids are synthesized in the rough endoplasmic reticulum and are then transferred to other cellular membranes. A sequence of transfer steps suggested by the order of increases in labeling of different types of membranes is rough endoplasmic reticulum, smooth membranes and nuclear membranes, vacuoles, and the plasma membrane.

Key words

Phospholipids Membranes Radioautography Amoebae 
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References

  1. Bennett G, Leblond CP, Haddad A (1974) Migration of glycoprotein from the Golgi apparatus to the surface of various cell types as shown by radioautography after labeled fucose injection into rats. J Cell Biol 60:258–284Google Scholar
  2. Bergmann JE, Tokuyasu KT, Singer SJ (1981) Passage of an integral membrane protein, the vesicular stomatitis virus glycoprotein, through the Golgi apparatus en route to the plasma membrane. Proc Natl Acad Sci USA 78:1746–1750Google Scholar
  3. Chlapowski FJ, Band RN (1971) Assembly of lipids into membranes in Acanthamoeba palestinensis. I. Observation on the specificity and stability of choline-14C and glycerol-3H as labels for membrane phospholipids. II. The origin and fate of glycerol-3H-labeled phospholipids of cellular membranes. J Cell Biol 50:625–633, 634–651Google Scholar
  4. Dallner G, Siekevitz P, Palade GE (1966) Biogenesis of endoplasmic reticulum membranes. I. Structural and chemical differentiation in developing rat hepatocyte. J Cell Biol 30:73–96Google Scholar
  5. Daniels EW (1973) Ultrastructure. In: Jeon KW (ed) The biology of amoeba. Academic Press, New York, pp 125–169Google Scholar
  6. Fawcett DW (1965) Structural and functional variations in the membranes of the cytoplasm. In: Seno S, Cowdry EV (eds) Intracellular membranous structure. Japan Society for Cell Biology, Okayama, pp 15–40Google Scholar
  7. Flickinger CJ (1969) The development of Golgi complexes and their dependence upon the nucleus in amebae. J Cell Biol 43:250–262Google Scholar
  8. Flickinger CJ (1973) Cellular membranes of amoeba. In: Jeon KW (ed) The biology of amoeba. Academic Press, New York, pp 171–199Google Scholar
  9. Flickinger CJ (1974a) Radioactive labeling of the Golgi apparatus by micro-injection of individual amebae. Exp Cell Res 88:415–418Google Scholar
  10. Flickinger CJ (1974b) Synthesis, intracellular transport, and release of secretory protein in the seminal vesicle of the rat, as studied by electron microscope radioautography. Anat Rec 180:407–426Google Scholar
  11. Flickinger CJ (1974) The fine structure of four “species” of Amoeba. J Protozool 21:59–68Google Scholar
  12. Flickinger CJ (1975) The relation between the Golgi apparatus, cell surface, and cytoplasmic vesicles in amoebae studied by electron microscope radioautography. Exp Cell Res 96:189–201Google Scholar
  13. Higgins JA (1974) Studies on the biogenesis of smooth endoplasmic reticulum membranes in hepatocytes of phenobarbital-treated rats. II. The site of phospholipid synthesis in the initial phase of membrane proliferation. J Cell Biol 62:635–646Google Scholar
  14. Higgins JA, Barrnett RJ (1972) Studies on the biogenesis of smooth endoplasmic reticulum membranes in livers of phenobarbital-treated rats. I. The site of activity of acyl-transferases involved in synthesis of the membrane phospholipid. J Cell Biol 55:282–298Google Scholar
  15. Ito S (1969) Structure and function of the glycocalyx. Fed Proc 28:12–25Google Scholar
  16. Jeon KW (1970) Micromanipulation of amoeba nuclei. Methods Cell Physiol 4:179–194Google Scholar
  17. Jeon KW, Lorch IJ (1973) Strain specificity in Amoeba proteus. In: Jeon KW (ed) The biology of amoeba. Academic Press, New York, pp 549–567Google Scholar
  18. Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 27:137AGoogle Scholar
  19. Maruta H, Goldstein L (1975) The fate and origin of the nuclear envelope during and after mitosis in Amoeba proteus. I. Synthesis and behavior of phospholipids of the nuclear envelope during the cell life cycle. J Cell Biol 65:631–645Google Scholar
  20. Michaels JE, Leblond CP (1976) Transport of glycoprotein from Golgi apparatus to cell surface by means of “carrier” vesicles as shown by radioautography of mouse colonic epithelium after injection of 3H-fucose. J Microscopie 25:243–247Google Scholar
  21. Morré DJ, Keenan TW, Huang CM (1974) Membrane flow and differentiation: Origin of Golgi apparatus membranes from endoplasmic reticulum. Adv Cytopharm 2:107–125Google Scholar
  22. Palade G (1975) Intracellular aspects of protein synthesis. Science 189:347–358PubMedGoogle Scholar
  23. Prescott DM (1964) Autoradiography with liquid emulsion. Methods Cell Physiol 1:365–370Google Scholar
  24. Prescott DM, Carrier RF (1964) Experimental procedures and cultural methods for Euplotes eurystomus and Ameba proteus. Methods Cell Physiol 1:85–95Google Scholar
  25. Read GA, Flickinger CJ (1980) Changes in production and turnover of surface components labeled with [3H] mannose in amoebae exposed to a general anesthetic. Exptl Cell Res 127:115–126Google Scholar
  26. Rothman JE, Fries E (1981) Transport of newly synthesized vesicular stomatitis viral glycoprotein to purified Golgi membranes. J Cell Biol 89:162–168Google Scholar
  27. Salpeter MM, McHenry FA (1973) Electron microscope autoradiography. Analyses of autoradiograms. In: Koehler JK (ed) Advanced techniques in biological electron microscopy. Springer-Verlag, Berlin, pp 113–152Google Scholar
  28. Stein O, Stein Y (1967) Lipid synthesis, intracellular transport, storage, and secretion. I. Electron microscopic radioautographic study of liver after injection of tritiated palmitate or glycerol in fasted and ethanol-treated rats. J Cell Biol 33:319–339Google Scholar
  29. Stein O, Stein Y (1969) Lecithin synthesis, intracellular transport, and secretion in rat liver. IV. A radioautographic and biochemical study of choline-deficient rats injected with choline-3H. J Cell Biol 40:461–483Google Scholar
  30. Stevens AR (1966) High resolution autoradiography. Methods Cell Physiol 2:255–310Google Scholar
  31. Sturgess J, Moscarello M, Schachter H (1978) The structure and biosynthesis of membrane glycoproteins. Curr Top Membrane Trans 11:16–105Google Scholar
  32. Wilgram GF, Kennedy EP (1963) Intracellular distribution of some enzymes catalyzing reactions in the biosynthesis of complex lipids. J Biol Chem 238:2615–2619Google Scholar
  33. Zar JH (1974) Biostatistical analysis. Prentice-Hall, Inc, Englewood Cliffs, New JerseyGoogle Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • Charles J. Flickinger
    • 1
  • Graham A. Read
    • 1
  1. 1.Department of AnatomyUniversity of Virginia, School of MedicineCharlottesvilleUSA

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