Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1979 Jul 1;82(1):114–139. doi: 10.1083/jcb.82.1.114

Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality

PMCID: PMC2110423  PMID: 479294

Abstract

The cytoplasmic ground substance of cultured cells prepared for high voltage transmission electron microscopy (glutaraldehyde/osmium fixed, alcohol or acetone dehydrated, critical-point dried) consists of slender (3-6 nm Diam) strands--the microtrabeculae (55)--that form an irregular three-dimensional lattice (the microtrabecular lattice). The microtrabeculae interconnect the membranous and nonmembranous organelles and are confluent with the cortices of the cytoplast. The lattice is found in all portions of the cytoplast of all cultured cells examined. The possibility that the lattice structure is an artifact of specimen preparation has been tested by (a) subjecting whole cultured cells (WI-38, NRK, chick embryo fibroblasts) to various chemical (aldehydes, osmium tetroxide) and nonchemical (freezing) fixation schedules, (b) examination of model systems (erythrocytes, protein solutions), (c) substantiating the relaibility of critical-point drying, and (d) comparing images of whole cells with conventionally prepared (plastic-embedded) cells. The lattice structure is preserved by chemical and nonchemical fixation, though alterations in ultrastructure can occur especially after prolonged exposure to osmium tetroxide. The critical-point method for drying specimens appears to be reliable as is the freeze-drying method. The discrepancies between images of plastic-embedded and sectioned cells, and images of whole, critical-point dried cells appear to be related, in part, to the electron-scattering properties of the embedding resin. The described observations indicate that the microtrabecular lattice seen in electron micrographs closely represents the nonrandom structure of the cytoplasmic ground substance of living cultured cells.

Full Text

The Full Text of this article is available as a PDF (2.9 MB).

Selected References

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

  1. Aaronson S. A., Weaver C. A. Characterization of murine sarcoma virus (Kirsten) transformation of mouse and human cells. J Gen Virol. 1971 Nov;13(2):245–252. doi: 10.1099/0022-1317-13-2-245. [DOI] [PubMed] [Google Scholar]
  2. Bloodgood R. A., Miller K. R. Freeze-fracture of microtubules and bridges in motile axostyles. J Cell Biol. 1974 Sep;62(3):660–671. doi: 10.1083/jcb.62.3.660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buckley I. K., Porter K. R. Cytoplasmic fibrils in living cultured cells. A light and electron microscope study. Protoplasma. 1967;64(4):349–380. doi: 10.1007/BF01666538. [DOI] [PubMed] [Google Scholar]
  4. Buckley I. K., Porter K. R. Electron microscopy of critical point dried whole cultured cells. J Microsc. 1975 Jul;104(2):107–120. doi: 10.1111/j.1365-2818.1975.tb04010.x. [DOI] [PubMed] [Google Scholar]
  5. Buckley I. K. Three dimensional fine structure of cultured cells: possible implications for subcellular motility. Tissue Cell. 1975;7(1):51–72. doi: 10.1016/s0040-8166(75)80007-3. [DOI] [PubMed] [Google Scholar]
  6. Bunge M. B. Fine structure of nerve fibers and growth cones of isolated sympathetic neurons in culture. J Cell Biol. 1973 Mar;56(3):713–735. doi: 10.1083/jcb.56.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burton P. R., Fernandez H. L. Delineation by lanthanum staining of filamentous elements associated with the surfaces of axonal microtubules. J Cell Sci. 1973 Mar;12(2):567–583. doi: 10.1242/jcs.12.2.567. [DOI] [PubMed] [Google Scholar]
  8. Byers H. R., Porter K. R. Transformations in the structure of the cytoplasmic ground substance in erythrophores during pigment aggregation and dispersion. I. A study using whole-cell preparations in stereo high voltage electron microscopy. J Cell Biol. 1977 Nov;75(2 Pt 1):541–558. doi: 10.1083/jcb.75.2.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chang J. H. Fixation and embedment, in situ, of tissue culture cells for electron microscopy. Tissue Cell. 1972;4(4):561–574. doi: 10.1016/s0040-8166(72)80030-2. [DOI] [PubMed] [Google Scholar]
  10. Ellisman M. H., Rash J. E., Staehelin L. A., Porter K. R. Studies of excitable membranes. II. A comparison of specializations at neuromuscular junctions and nonjunctional sarcolemmas of mammalian fast and slow twitch muscle fibers. J Cell Biol. 1976 Mar;68(3):752–774. doi: 10.1083/jcb.68.3.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldman R. D. The use of heavy meromyosin binding as an ultrastructural cytochemical method for localizing and determining the possible functions of actin-like microfilaments in nonmuscle cells. J Histochem Cytochem. 1975 Jul;23(7):529–542. doi: 10.1177/23.7.1095652. [DOI] [PubMed] [Google Scholar]
  12. Gray E. G. Synaptic fine structure and nuclear, cytoplasmic and extracellular networks: The stereoframework concept. J Neurocytol. 1975 Jun;4(3):315–339. doi: 10.1007/BF01102116. [DOI] [PubMed] [Google Scholar]
  13. Hinkley R. E., Jr Axonal microtubules and associated filaments stained by Alcian blue. J Cell Sci. 1973 Nov;13(3):753–761. doi: 10.1242/jcs.13.3.753. [DOI] [PubMed] [Google Scholar]
  14. Ishikawa H., Bischoff R., Holtzer H. Formation of arrowhead complexes with heavy meromyosin in a variety of cell types. J Cell Biol. 1969 Nov;43(2):312–328. [PMC free article] [PubMed] [Google Scholar]
  15. LeBeux Y. J., Willemot J. An ultrastructural study of the microfilaments in rat brain by means of E-PTA staining and heavy meromyosin labeling. II. The synapses. Cell Tissue Res. 1975 Jun 27;160(1):37–68. doi: 10.1007/BF00219841. [DOI] [PubMed] [Google Scholar]
  16. LeBeux Y. J., Willemot J. An ultrastructural study of the microfilaments in rat brain by means of heavy meromyosin labeling. I. The perikaryon, the dendrites and the axon. Cell Tissue Res. 1975 Jun 27;160(1):1–36. doi: 10.1007/BF00219840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lenk R., Ransom L., Kaufmann Y., Penman S. A cytoskeletal structure with associated polyribosomes obtained from HeLa cells. Cell. 1977 Jan;10(1):67–78. doi: 10.1016/0092-8674(77)90141-6. [DOI] [PubMed] [Google Scholar]
  18. Mazia D., Schatten G., Sale W. Adhesion of cells to surfaces coated with polylysine. Applications to electron microscopy. J Cell Biol. 1975 Jul;66(1):198–200. doi: 10.1083/jcb.66.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McNutt N. S. A thin-section and freeze-fracture study of microfilament-membrane attachments in choroid plexus and intestinal microvilli. J Cell Biol. 1978 Dec;79(3):774–787. doi: 10.1083/jcb.79.3.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mooseker M. S., Tilney L. G. Organization of an actin filament-membrane complex. Filament polarity and membrane attachment in the microvilli of intestinal epithelial cells. J Cell Biol. 1975 Dec;67(3):725–743. doi: 10.1083/jcb.67.3.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. PALADE G. E. A study of fixation for electron microscopy. J Exp Med. 1952 Mar;95(3):285–298. doi: 10.1084/jem.95.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. PALADE G. E., SIEKEVITZ P. Liver microsomes; an integrated morphological and biochemical study. J Biophys Biochem Cytol. 1956 Mar 25;2(2):171–200. doi: 10.1083/jcb.2.2.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. PORTER K. R. Observations on a submicroscopic basophilic component of cytoplasm. J Exp Med. 1953 May;97(5):727–750. doi: 10.1084/jem.97.5.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reaven E. P., Axline S. G. Subplasmalemmal microfilaments and microtubules in resting and phagocytizing cultivated macrophages. J Cell Biol. 1973 Oct;59(1):12–27. doi: 10.1083/jcb.59.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. SABATINI D. D., BENSCH K., BARRNETT R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. doi: 10.1083/jcb.17.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sale W. S., Satir P. Splayed Tetrahymena cilia. A system for analyzing sliding and axonemal spoke arrangements. J Cell Biol. 1976 Nov;71(2):589–605. doi: 10.1083/jcb.71.2.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Smith D. S., Järlfors U., Beránek R. The organization of synaptic axcplasm in the lamprey (petromyzon marinus) central nervous system. J Cell Biol. 1970 Aug;46(2):199–219. doi: 10.1083/jcb.46.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Spooner B. S., Yamada K. M., Wessells N. K. Microfilaments and cell locomotion. J Cell Biol. 1971 Jun;49(3):595–613. doi: 10.1083/jcb.49.3.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tani E., Ametani T. Substructure of microtubules in brain nerve cells as revealed by ruthenium red. J Cell Biol. 1970 Jul;46(1):159–165. doi: 10.1083/jcb.46.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tilney L. G. How microtubule patterns are generated. The relative importance of nucleation and bridging of microtubules in the formation of the axoneme of Raphidiophrys. J Cell Biol. 1971 Dec;51(3):837–854. doi: 10.1083/jcb.51.3.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vial J., Porter K. R. Scanning microscopy of dissociated tissue cells. J Cell Biol. 1975 Nov;67(2PT1):345–360. doi: 10.1083/jcb.67.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wessells N. K., Spooner B. S., Ludueña M. A. Surface movements, microfilaments and cell locomotion. Ciba Found Symp. 1973;14:53–82. doi: 10.1002/9780470719978.ch4. [DOI] [PubMed] [Google Scholar]
  33. Wolosewick J. J., Porter K. R. Observation on the morphological heterogeneity of WI-38 cells. Am J Anat. 1977 Jun;149(2):197–225. doi: 10.1002/aja.1001490206. [DOI] [PubMed] [Google Scholar]
  34. Wolosewick J. J., Porter K. R. Stereo high-voltage electron microscopy of whole cells of the human diploid line, WI-38. Am J Anat. 1976 Nov;147(3):303–323. doi: 10.1002/aja.1001470305. [DOI] [PubMed] [Google Scholar]
  35. Yamada K. M., Spooner B. S., Wessells N. K. Ultrastructure and function of growth cones and axons of cultured nerve cells. J Cell Biol. 1971 Jun;49(3):614–635. doi: 10.1083/jcb.49.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES