Skip to main content

Normal and pseudorabies virus infected primary nerve cell cultures in scanning electron microscopy

Medical Microbiology and Immunology volume 167, pages 117–126 (1979)Cite this article

Abstract

Primary cell cultures from the central nervous system of the embryonic rat were inoculated with pseudorabies virus. Their morphological changes were studied by phase contrast microscopy and by scanning as well as by transmission electron microscopy. Uninfected cultures display two distinct cell layers in scanning electron microscopy: a flat continuous monolayer supports a heterogeneous population of individual, presumably neural cells, which emit processes of different number and size. The latter cells form contacts by a dense network of fibres.

Infectious virus is propagated in these nerve cell cultures with similar effectivity as in other cultures. The infection leads to fusion and death of the cells. By the time the cytopathic effect is visible, nearly all cells, including those of neuronal and those of nonneuronal appearance, are studded with ample amounts of virus-sized particles. The particles represent viruses as demonstrated by transmission electron microscopy or by treatment with a hyperimmune serum directed against pseudorabies virus structural components. Hyperimmune serum leads to clustering of the particles at the cell surface. The amount of virus particles per surface unit was about 10 times higher on neural cells as compared to primary rabbit kidney cells. The concentration of infectious particles in the supernatant, however was approximately the same. The system described appears to be useful for the study of acute virus effects on neural tissue under strictly controlled conditions.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  • Darlington, R.W., Granoff, A.: Replication Biological Aspects. In: A.S. Kaplan (ed.) The Herpes viruses. pp. 93–129. New York: Acad. Press 1973

    Google Scholar 

  • Dempsher, J., Larrabee, M. G., Bang, F. B., Bodian, D.: Physiological changes in sympathetic ganglia infected with Pseudorabies virus. Am. J. Physiol.182, 203–216 (1955)

    PubMed  Google Scholar 

  • Dimpfel, W., Habermann, E.: Binding characteristics of125I-labelled tetanus toxin to primary tissue cultures from mouse embryonic CNS. J. Neurochem.29, 1111–1120 (1977)

    PubMed  Google Scholar 

  • Dimpfel, W., Huang, R. T. C., Habermann, E.: Gangliosides in nervous tissue cultures and binding of125I-labelled tetanus toxin, a neuronal marker. J. Neurochem.29, 329–334 (1977)

    PubMed  Google Scholar 

  • Dolivo, M., Beretta, E., Bonifas, V., Foroglou, C.: Ultrastructure and function in sympathetic ganglia isolated from rats infected with Pseudorabies virus. Brain Res.140, 111–123 (1978)

    PubMed  Google Scholar 

  • Echlin, P.: Sputter coating techniques for scanning electron microscopy. Scanning Electron Microscopy Part I, 217–232 (1975)

  • Felluga, B.: Electron microscopic observations on Pseudorabies virus development in a line of pig kidney cells. Ann. Sclavo5, 412–424 (1963)

    Google Scholar 

  • Fonte, V. G., Porter, K. R.: Visualization in whole cells of Herpes simplex virus using SEM and TEM. Scanning electron microscopy Part III, 827–834 (1974)

  • Giller, E. L., Schrier, B. K., Shainberg, A., Fisk, H. R., Nelson, P. G.: Choline acetyltransferase activity is increased in combined cultures of spinal cord and muscle cells from mice. Science182, 588–589 (1973)

    PubMed  Google Scholar 

  • Gilman, A. G., Schrier, B. K.: Adenosine cyclic 3′5′-monophosphate in fetal rat brain cell cultures. I. Effect of catecholamines. Molec. Pharmacol.8, 410–416 (1972)

    Google Scholar 

  • Godfrey, E. W., Nelson, P. G., Schrier, B. K., Breuer, A. C., Ransom, B. R.: Neurons from fetal rat brain in a new cell culture system: A multidisciplinary analysis. Brain Res.90, 1–21 (1975)

    PubMed  Google Scholar 

  • Kaplan, A. S. Herpes simplex and Pseudorabies viruses. Virology Monographs5 (1969)

  • Ludwig, H., Becht, H., Rott, R.: Inhibition of Herpes virus-induced cell fusion by concanavalin A, antisera, and 2-deoxy-D-glucose. J. Virol.14, 307–314 (1974)

    PubMed  Google Scholar 

  • Morgan, C., Rose, H. M., Mednis, B.: Electron microscopy of Herpes simplex virus. J. Virol.2, 507–516 (1968)

    PubMed  Google Scholar 

  • Nii, S., Morgan, C., Rose, H. M.: Electron microscopy of Herpes simplex virus. J. Virol.2, 517–536 (1968)

    PubMed  Google Scholar 

  • Noel-Courtey, B., Heinen, E.: Observation au microscope électronique à balayage de cellules nerveuses de moelle spinale d'embryons de poulet cultivées in vitro sur la polylysine-L. O. C. R. Acad. Sci. Paris285, Serie D, 385–387 (1977)

    Google Scholar 

  • Peacock, J. H., Nelson, P. G., Goldstone, M. W.: Electrophysiologic study of cultured neurons dissociated from spinal cords and dorsal root ganglia of fetal mice. Dev. Biol.30, 137–153 (1973)

    PubMed  Google Scholar 

  • Poste, G., Allison, A. C.: Membrane fusion. Biochim. Biophys. Acta300, 421–465 (1973)

    PubMed  Google Scholar 

  • Ransom, B. R., Neale, E., Henkart, M., Bullock, P. N., Nelson P. G.: Mouse spinal cord in cell cultures. I. Morphology and intrinsic neuronal electrophysiological properties. J. Neurophysiol.40, 1132–1150 (1977)

    PubMed  Google Scholar 

  • Rodriguez, M., Dubois-Dalcq, M.: Intramembrane changes occurring during maturation of Herpes simplex virus type 1: Freeze-fracture study. J. Virol.26, 435–447 (1978)

    PubMed  Google Scholar 

  • Schlehofer, J. R., Habermehl, K.-O.: Herpes simplex virus induced alterations of membrane morphology and permeability of cells. 9th Intern. Congr. Electr. Microscopy Vol. II, 374–375 (1978)

    Google Scholar 

  • Schrier, B. K.: Surface culture of fetal mammalian brain cells: effect of subculture on morphology and choline acetyltransferase activity. J. Neurobiol.4, 117–124 (1973)

    PubMed  Google Scholar 

  • Schrier, B. K., Shapiro, D. L.: Effects of fluorodeoxyuridine on growth and choline acetyltransferase activity in fetal rat brain cells in surface culture. J. Neurobiol.5 151–159 (1974)

    PubMed  Google Scholar 

  • Shimada, Y., Fishman, D. A.: Scanning electron microscopy of nerve muscle contacts in embryonic cell culture. Devel. Biol.43, 42–61 (1975)

    Google Scholar 

Download references

Author information

Author notes

  1. H. Ludwig

    Present address: Institut für Virologie, Freie UniversitÄt Berlin, Deutschland

Affiliations

  1. Pharmakologisches Institut (FB 23) der Justus Liebig-UniversitÄt Giessen, Deutschland

    U. Bijok, W. Dimpfel & E. Habermann

  2. Institut für Virologie (FB 18) der Justus Liebig-UniversitÄt Giessen, Frankfurter Strasse 107, D-6300, Lahn-Giessen 1, Federal Republic of Germany

    H. Ludwig

Authors
  1. U. Bijok
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. W. Dimpfel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E. Habermann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H. Ludwig
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Part of this work will be presented in the thesis of U. Bijok

Work was supported by Sonderforschungsbereich 47 and the Bundesministerium des Inneren.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bijok, U., Dimpfel, W., Habermann, E. et al. Normal and pseudorabies virus infected primary nerve cell cultures in scanning electron microscopy. Med Microbiol Immunol 167, 117–126 (1979). https://doi.org/10.1007/BF02123561

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02123561

Keywords

  • Nerve Cell Culture
  • Primary Cell Culture
  • Primary Nerve Cell
  • Primary Nerve Cell Culture
  • Rabbit Kidney