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. 1978 Sep 1;78(3):756–768. doi: 10.1083/jcb.78.3.756

Freeze-fracture studies of the developing cell surface. II. Particle- free membrane blisters on glutaraldehyde-fixed corneal fibroblasts are artefacts

PMCID: PMC2110182  PMID: 100501

Abstract

We describe, in sections and by freeze-fracture, four classes of intramembrane particle (IMP)-free membrane blebs or "blisters" associated with glutaraldehyde-fixed embryonic corneal fibroblasts: (a) Single blisters attached to the cell membrane; (b) free (detached) vesicles; (c) myelin figures; (d) multivesicular protrusions which resemble the "mounds" described by others on nerve growth cones. The IMP-free, membrane-bounded blisters contain no ground cytoplasm or organelles, in contrast to blebs on trypsin-isolated fibroblasts, which we show here do contain cytoplasm and IMP-rich membranes. That the IMP- free membrane blisters in embryonic corneas are artefacts of fixation is demonstrated by (a) their absence in replicas of fibroblasts frozen and fractured without prior aldehyde fixation and (b) their absence in sections of fibroblasts fixed in a combination of glutaraldehyde and osmium tetroxide. We suggest that the addition of osmium prevents postfixation movement of membrane lipids, especially the negatively charged "fluid" lipids which others have shown are capable of considerable mobility after aldehyde fixation alone. Recent literature has implicated membrane blistering in secretory processes and in growth of nerves, but before the functional significance of such IMP-free blisters is assessed, membrane mobility of the type shown here should be taken into consideration.

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Selected References

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  1. Arborgh B., Bell P., Brunk U., Collins V. P. The osmotic effect of glutaraldehyde during fixation. A transmission electron microscopy, scanning electron microscopy and cytochemical study. J Ultrastruct Res. 1976 Sep;56(3):339–350. doi: 10.1016/s0022-5320(76)90009-5. [DOI] [PubMed] [Google Scholar]
  2. Bard J. B., Hay E. D., Meller S. M. Formation of the endothelium of the avian cornea: a study of cell movement in vivo. Dev Biol. 1975 Feb;42(2):334–361. doi: 10.1016/0012-1606(75)90339-5. [DOI] [PubMed] [Google Scholar]
  3. Bloom G. D., Haegermark O. A study on morphological changes and histamine release induced by compound 48/80 in rat peritoneal mast cells. Exp Cell Res. 1965 Dec;40(3):637–654. doi: 10.1016/0014-4827(65)90241-7. [DOI] [PubMed] [Google Scholar]
  4. Bunge M. B. Initial endocytosis of perioxidase or ferritin by growth cones of cultured nerve cells. J Neurocytol. 1977 Aug;6(4):407–439. doi: 10.1007/BF01178226. [DOI] [PubMed] [Google Scholar]
  5. Chi E. Y., Lagunoff D., Koehler J. K. Freeze-fracture study of mast cell secretion. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2823–2827. doi: 10.1073/pnas.73.8.2823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hasty D. L., Hay E. D. Freeze-fracture studies of the developing cell surface. I. The plasmalemma of the corneal fibroblast. J Cell Biol. 1977 Mar;72(3):667–686. doi: 10.1083/jcb.72.3.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Heuser J. E., Reese T. S., Landis D. M. Preservation of synaptic structure by rapid freezing. Cold Spring Harb Symp Quant Biol. 1976;40:17–24. doi: 10.1101/sqb.1976.040.01.004. [DOI] [PubMed] [Google Scholar]
  8. Hirsch J. G., Fedorko M. E. Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and "postfixation" in uranyl acetate. J Cell Biol. 1968 Sep;38(3):615–627. doi: 10.1083/jcb.38.3.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Land B. R., Podleski T. R., Salpeter E. E., Salpeter M. M. Acetylcholine receptor distribution on myotubes in culture correlated to acetylcholine sensitivity. J Physiol. 1977 Jul;269(1):155–176. doi: 10.1113/jphysiol.1977.sp011897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lawson D., Raff M. C., Gomperts B., Fewtrell C., Gilula N. B. Molecular events during membrane fusion. A study of exocytosis in rat peritoneal mast cells. J Cell Biol. 1977 Feb;72(2):242–259. doi: 10.1083/jcb.72.2.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nermut M. V., Ward B. J. Effect of fixatives on fracture plane in red blood cells. J Microsc. 1974 Sep;102(Pt 1):29–39. doi: 10.1111/j.1365-2818.1974.tb03963.x. [DOI] [PubMed] [Google Scholar]
  12. PETERS A. The structure of myelin sheaths in the central nervous system of Xenopus laevis (Daudin). J Biophys Biochem Cytol. 1960 Feb;7:121–126. doi: 10.1083/jcb.7.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pfenninger K. H., Bunge R. P. Freeze-fracturing of nerve growth cones and young fibers. A study of developing plasma membrane. J Cell Biol. 1974 Oct;63(1):180–196. doi: 10.1083/jcb.63.1.180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. REVEL J. P., ITO S., FAWCETT D. W. Electron micrographs of myelin figures of phospholipide simulating intracellular membranes. J Biophys Biochem Cytol. 1958 Jul 25;4(4):495–498. doi: 10.1083/jcb.4.4.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schneeberger E. E., Lynch R. D., Geyer R. P. Glutaraldehyde fixation used to demonstrate altered properties of outer mitochondrial membranes in polyunsaturated fatty acid (PUFA) supplemented cells. Exp Cell Res. 1976 Jun;100(1):117–128. doi: 10.1016/0014-4827(76)90334-7. [DOI] [PubMed] [Google Scholar]
  16. Toister Z., Loyter A. The mechanism of cell fusion. II. Formation of chicken erythrocyte polykaryons. J Biol Chem. 1973 Jan 25;248(2):422–432. [PubMed] [Google Scholar]
  17. Trump B. F., Bulger R. E. New ultrastructural characteristics of cells fixed in a glutaraldehyde-osmium tetroxide mixture. Lab Invest. 1966 Jan;15(1 Pt 2):368–379. [PubMed] [Google Scholar]
  18. da Silva P. P., Miller R. G. Membrane particles on fracture faces of frozen myelin. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4046–4050. doi: 10.1073/pnas.72.10.4046. [DOI] [PMC free article] [PubMed] [Google Scholar]

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