Advertisement

Virchows Archiv B

, Volume 29, Issue 1, pp 281–296 | Cite as

Studies on cell surface conformation following injury

I. Scanning and transmission electron microscopy of cell surface changes following P-chloromercuribenzene sulfonic acid (PCMBS)-induced injury of ehrlich ascites tumor cells
  • Benjamin F. Trump
  • Antti Penttila
  • Irene K. Berezesky
Article

Summary

In the present series of studies, the use of scanning electron microscopy combined with analysis of thin sections was applied to the examination of cell membrane changes following acute lethal cell injury. In this report, acute injury of Ehrlich ascites tumor cells directed primarily at the plasma membrane was produced with the non-penetrating mercurial p-chloromercuribenzene sulfonate (PCMBS). This compound interacts initially with the cell membrane, primarily by modifying membrane associated sulfhydryl groups resulting in rapid changes in membrane permeability. The present morphological changes revealed that rapid swelling and ultimately lysis of the cells occurred. These have earlier been shown to be accompanied by marked ion and water shifts and a concomitant decrease in adenine nucleotide content. Changes in cell surface contours as seen by scanning electron microscopy were dramatic and included rapid loss of normal surface microvillous patterns, and formation of surface folds, broad-based elevations and blebs with numerous intricate recesses. In transmission electron microscopy, membrane-limited vacuoles were seen at the cell periphery in addition to the above changes. The present study emphasizes the lability of cell surface conformation toward this toxic non-penetrating mercurial compound and the important role of cell membrane changes in acute lethal cell injury.

Key words

Cell surface changes EATC p-chloromercuribenzene sulfonic acid-indiced injury 

References

  1. Anderson, T.F.: Techniques for the preservation of three-dimensional structure in preparing specimens for electron microscope. Trans. NY Acad. Sci.13, 130–134 (1951)Google Scholar
  2. Arstila, A.U., Jauregui, H.O., Chang, J., Trump, B.F.: Studies on cellular autophagocytosis. Relationship between heterophagy and autophagy in HeLa cells. Lab. Invest.24, 162–174 (1971)PubMedGoogle Scholar
  3. Belkin, M., Hardy, W.G.: Relation between water permeability and integrity of sulfhydryl groups in malignant and normal cells. J. Biophys. Biochem. Cytol.9, 733–738 (1961)CrossRefPubMedGoogle Scholar
  4. Bessis, M., Weed, R.I.: The structure of normal and pathologic erythrocytes. Biol. Med. Physics14, 36–92 (1973)Google Scholar
  5. Boyde, A., Weiss, R.A., Vesely, P.: Scanning electron microscopy of cells in culture. Exp. Cell Res.71, 313–324(1972)PubMedCrossRefGoogle Scholar
  6. Carr, K.E.: Applications of scanning electron microscopy in biology. Int. Rev. Cytol.30, 183–255 (1971)PubMedCrossRefGoogle Scholar
  7. Cook, J.S.: Photopathology of the erythrocyte membrane. In: Pathobiology of cell membranes, (B.F. Trump and A.U. Arstila, eds.) Vol. I, pp. 199–216. New York: Academic Press 1975Google Scholar
  8. Costero, I., Pomerat, C.M.: Cultivation of neurons from the adult human cerebral and cerebellar cortex. Am. J. Anat.89, 405–468 (1951)PubMedCrossRefGoogle Scholar
  9. Estensen, R.: Toward understanding the mode of cytochalasin B. Action and associated cell functions. In: Molecular pathology, (R.A. Good, S.B. Day and J.J. Yunis, eds.), pp. 379–393. Springfield, IL: Charles C. Thomas 1975Google Scholar
  10. Follett, E.A.C., Goldman, R.D.: The occurrence of microvilli during spreading and growth of BHK21/C13 fibroblasts. Exp. Cell Res.59, 124–136 (1970)PubMedCrossRefGoogle Scholar
  11. Fuller, G.M., Ellison, J.J., McGill, M., Sordahl, L.A., Brinkley, B.R.: Studies on the inhibitory role of calcium in the regulation of microtubule assembly in vitro and in vivo. In: Microtubules and microtubule inhibitors, (M. Borgers and M. DeBrabander, eds.), pp. 379–390. Amsterdam: North-Holland Publishing Co. 1975Google Scholar
  12. Goldman, M.A., Leif, R.C.: A wet chemical method for rendering scanning electron microscopy samples conductive and observations on the surface morphology of human erythrocytes and Ehrlich ascites cells. Proc. Natl. Acad. Sci. U.S.A.70, 3599–3603 (1973)PubMedCrossRefGoogle Scholar
  13. Herdson, P.B., Kaltenbach, J.P.: Fine structural changes in hypotonically treated Ehrlich ascites tumor cells. Exp. Cell Res.42, 74–82 (1966)PubMedCrossRefGoogle Scholar
  14. Hsie, A.W., Puck, T.T.: Morphologic transformation of Chinese hamster cells by dibutyryl adenosine cyclic 3′-5′-monophosphate and testosterone. Proc. Natl. Acad. Sci. U.S.A.68, 358–361 (1971)PubMedCrossRefGoogle Scholar
  15. Kaltenbach, J.P., Kaltenbach, M.H., Lyons, W.B.: Nigrosin as a dye for differentiating live and dead ascites cells. Exp. Cell Res.15, 112–117 (1958)PubMedCrossRefGoogle Scholar
  16. Kelley, R.O., Dekker, R.A.F., Bluemink, J.G.: Ligand-mediated osmium binding: Its application in coating biological specimens for scanning electron microscopy. J. Ultrastr. Res.45, 254–258 (1973)CrossRefGoogle Scholar
  17. King, D.W., Paulson, S.R., Hannaford, N.C., Krebs, A.T.: Cell death. I. The effect of injury on the proteins and deoxyribonucleic acid of Ehrlich ascites tumor cells. Am. J. Path.35, 369–382 (1959)PubMedGoogle Scholar
  18. Laiho, K.U., Shelburne, J.D., Trump, B.F.: Observations on cell volume, ultrastructure, mitochondrial conformation and vital-dye uptake in Ehrlich ascites tumor cells. Am. J. Path.65, 203–230 (1971)PubMedGoogle Scholar
  19. Laiho, K.U., Trump, B.F.: Relationship of ionic, water, and cell volume changes in cellular injury of Ehrlich ascites tumor cells. Lab. Invest.31, 207–215 (1974a)PubMedGoogle Scholar
  20. Laiho, K.U., Trump, B.F.: The relationship between cell viability and changes in mitochondrial ultrastructure, cellular ATP, ion and water content following injury of Ehrlich ascites tumor cells. Virchows Arch. B Cell Path.15, 267–277 (1974 b)Google Scholar
  21. Lianes, A.T., Fare, C.B., Ferenczy, A., Richart, R.M.: Scanning electron microscopy of normal exfoliated squamous cervical cells. Acta Cytol.17, 507–509 (1973)Google Scholar
  22. Penttila, A., Trump, B.F.: Studies on modification of the cellular response to injury. I. Protective effect of acidosis on p-choloromercuribenzene sulfonic acid-induced injury of Ehrlich ascites tumor cells. Lab. Invest.32, 690–695 (1975a)PubMedGoogle Scholar
  23. Penttila, A., Trump, B.F.: Studies on the modification of the cellular response to injury. II. Electron microscopic studies on the protective effect of acidosis on anoxic injury of Ehrlich ascites tumor cells. Virchows Arch. B Cell Path.18, 1–16 (1975b)Google Scholar
  24. Penttila, A., Trump, B.F.: Studies on the modification of the cellular response to injury. III. Electron microscopic studies on the protective effect of acidosis on p-choloromercuribenzene sulfonic acid-(PCMBS) induced injury of Ehrlich ascites tumor cells. Virchows Arch. B Cell Path.18, 17–34 (1975 c)Google Scholar
  25. Perecko, J.P., Berezesky, I.K., Grimley, P.M.: Surface features of some established murine cell lines under various conditions of oncogenic virus infection. In: Scanning electron microscopy, (O. Johari and I. Corvin, eds.), pp. 521–527. Chicago, IL: IIT Research Institute 1973Google Scholar
  26. Polliack, A., McKenzie, S., Gee T., Lampen, N., DeHarven, E., Clarkson, B.D.: A scanning electron microscopic study of 34 cases of acute granulocytic, myelomonocytic, monoblastic and histiocytic leukemia. Am. J. Med.59, 308–315 (1975)PubMedCrossRefGoogle Scholar
  27. Porter, K.R., Fonte, V., Weiss, G.: A scanning microscope study of the topography of HeLa cells. Cancer Res.34, 1385–1394 (1974)PubMedGoogle Scholar
  28. Porter, K.R., Prescott, D., Frye, J.: Changes in surface morphology of Chinese hamster ovary cells during the cell cycle. J. Cell Biol.57, 815–836 (1973a)PubMedCrossRefGoogle Scholar
  29. Porter, K.R., Todaro, G.J., Fonte, V.: A scanning electron microscope study of surface features of viral and spontaneous transformants of mouse BALB/3T3 cells. J. Cell Biol.59, 633–642 (1973 b)PubMedCrossRefGoogle Scholar
  30. Porter, K.R., Weiss, G.: Chick fibroblasts during collagen production. A scanning electron microscope study. J. Cell Biol.57, 815–836 (1973)PubMedCrossRefGoogle Scholar
  31. Price, Z.H.: The micromorphology of zeiotic blebs in cultured human epithelial (HEp) cells. Exp. Cell Res.48, 82–99 (1967)PubMedCrossRefGoogle Scholar
  32. Puck, T.T., Waldren, C.A., Hsie, A.W.: Membrane dynamics in the action of dibutyryl adenosine 3′-5′-cyclic monophosphate and testosterone on mammalian cells. Proc. Nat’l. Acad. Sci. U.S.A.69, 1943–1947 (1972)CrossRefGoogle Scholar
  33. Pugh-Humphreys, R.G.P., Sinclair, W.: Ultrastructural studies relating to the surface morphology of cultured cells. J. Cell Sci.6, 477–484 (1970)PubMedGoogle Scholar
  34. Rothstein, A.: Sulfhydryl groups in red cell membranes. Exp. Eye Res.11, 329–337 (1971)PubMedCrossRefGoogle Scholar
  35. Rubin, R.W., Everhart, L.P.: The effect of cell-to-cell contact on the surface morphology of Chinese hamster ovary cells. J. Cell Biol.57, 837–844 (1973)PubMedCrossRefGoogle Scholar
  36. Sanders, S.K., Alexander, E.L., Braylan, R.C.: A high-yield technique for preparing cells fixed in suspension for scanning electron microscopy. J. Cell Biol.67, 476–480 (1975)PubMedCrossRefGoogle Scholar
  37. Shohet, S.B., Layzer, R.B.: The ‘muscle’ of the red cell. New Engl. J. Med.294, 221–222 (1976)PubMedGoogle Scholar
  38. Sugimura, M.J.: Scanning and transmission electron microscopy of individual cells. J. Electr. Micr.22, 367–369 (1973)Google Scholar
  39. Sutherland, R.M., Rothstein, A., Weed, R.I.: Erythrocyte membrane sulfhydryl groups and cation permeability. J. Cell Physiol.69, 185–198 (1967)PubMedCrossRefGoogle Scholar
  40. Trump, B.F., Croker, B.P., Mergner, W.J.: The role of energy metabolism, ion and water shifts in the pathogenesis of cell injury. In: Cell membranes: biological and pathologic aspects, (G.W. Richter and D.G. Scarpelli, eds.), pp. 84–128. Baltimore: Williams and Wilkins Co. 1971Google Scholar
  41. Trump, B.F., Ericsson, J.L.E.: Some ultrastructural and biochemical consequences of cell injury. In: The inflammatory process, (B.W. Zweibach and L. Grant, eds.), pp. 35–130. New York: Academic Press 1965Google Scholar
  42. Trump, B.F., Ginn, F.L.: Pathogenesis of subcellular reaction to lethal injury. In: Methods and achievements in experimental pathology, (E. Bajusz and G. Jasmin, eds.), Vol. 4, pp. 1–29. Basel and New York: Karger 1969Google Scholar
  43. Trump, B.F., Laiho, K.U., Mergner, W.J., Arstila, A.U.: Studies on the subcellular pathophysiology of acute lethal cell injury. Beitr. Path.152, 243–271 (1974)Google Scholar
  44. Trump, B.F., Penttila, A., Berezesky, I.K.: Studies on cell surface conformation following injury. II. Scanning and transmission electron microscopy of cell surface changes following anoxic injury in Ehrlich ascites tumor cells. Virchows Arch. B Cell Path.29, 297–307 (1979)Google Scholar
  45. Webb, J.L.: Enzyme and metabolic inhibitors, Vol. 2. New York: Academic Press 1966Google Scholar
  46. Weed, R.I., Reed, C.F.: Membrane alterations leading to red cell destruction. Am. J. Med.41, 681–698 (1966)PubMedCrossRefGoogle Scholar
  47. Wetzel, B., Cannon, G.B., Alexander, E.L., Erickson, B.W., Westbrook, E.W.: A critical approach to the scanning electron microscopy of cells in suspension. In: Scanning Electron Microscopy, (O. Johari and I. Corvin, eds.), pp. 581–588. Chicago, IL: IIT Research Institute 1973Google Scholar
  48. Zollinger, H.U.: Cytologie studies with the phase microscope. I. The formation of ‘blisters’ on cells in suspension (potocytosis), with observations on the nature of the cellular membrane. Am. J. Path.24, 545–568 (1948)PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1979

Authors and Affiliations

  • Benjamin F. Trump
    • 2
  • Antti Penttila
    • 1
    • 2
  • Irene K. Berezesky
    • 2
  1. 1.Department of Forensic MedicineUniversity of HelsinkiHelsinkiFinland
  2. 2.Department of PathologyUniversity of Maryland School of MedicineBaltimore

Personalised recommendations