1887

Abstract

SUMMARY

A rat phaeochromocytoma cell line, termed PC12, was used to study scrapie replication. These cells, in response to the addition of nerve growth factor (NGF), exhibit a number of neuronal properties including morphological differentiation, electrophysiological responsiveness, and neurotransmitter synthesis. Cultures were exposed to scrapie brain homogenate (strain 139A), harvested every week for up to 6 weeks, and assayed for scrapie infectivity. Scrapie replication was monitored by injecting scrapie agent-exposed NGF-treated PC12 cells into mice and measuring time intervals from injection to onset of clinical symptoms. Mouse incubation periods vary inversely with the amount of scrapie infectivity present. Cells harvested at 7 and 14 days after exposure to scrapie agent showed a decrease in the level of infectivity followed by an increase at subsequent time points. The increase in scrapie infectivity from early to late time intervals after agent exposure clearly indicated replication . A fusion agent was not necessary to establish infection, and the addition of mouse peritoneal macrophages caused a reduction in the yield of infectivity per culture. Examination of cells by phase-contrast microscopy failed to reveal any cytopathology.

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1984-12-01
2024-11-13
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References

  1. Bornstein M. B. 1958; Reconstituted rat tail collagen used as a substrate for tissue culture on coverslips in Maximow slides and roller tubes. Laboratory Investigation 1:134–137
    [Google Scholar]
  2. Buyukmihci N., Goehring-Harmon F., Marsh R. F. 1983; Neural pathogenesis of experimental scrapie after intraocular inoculation of hamsters. Experimental Neurology 81:396–406
    [Google Scholar]
  3. Carp R. I., Callahan S. M. 1981; In vitro interaction of scapie agent and mouse peritoneal macrophages. Intervirology 16:8–13
    [Google Scholar]
  4. Carp R. I., Callahan S. M. 1982; Effect of mouse peritoneal macrophages on scrapie infectivity during extended in vitro incubation. Intervirology 17:201–207
    [Google Scholar]
  5. Carp R. I., Callahan S. M., Sersen E. A., Moretz R. C. 1984; Preclinical changes in weight of scrapie-infected mice as a function of scrapie agent-mouse strain combination. Intervirology 21:61–69
    [Google Scholar]
  6. Clarke M. C. 1979; Infection of cell cultures with scrapie agent. In Slow Transmissible Diseases of the Nervous System vol 2: pp 225–233 Edited by Prusiner S. B., Hadlow W. J. New York: Academic Press;
    [Google Scholar]
  7. Clarke M. C., Haig D. A. 1970a; Evidence for the multiplication of scrapie agent in cell culture. Nature, London 225:100–101
    [Google Scholar]
  8. Clarke M. C., Haig D. A. 1970b; Multiplication of scrapie agent in cell culture. Research in Veterinary Science 11:500–501
    [Google Scholar]
  9. Clarke M. C., Millson G. C. 1976; Infection of a cell line of mouse L fibroblasts with scrapie agent. Nature, London 261:144–145
    [Google Scholar]
  10. Dickinson A. G., Fraser H. 1977; Scrapie: pathogenesis in inbred mice: an assessment of host control and response involving many strains of agent. In Slow Virus Infections of the Central Nervous System pp 3–14 Edited by Ter Meulen V., Katz M. New York: Springer-Verlag;
    [Google Scholar]
  11. Dickinson A. G., Outram G. W. 1979; The scrapie replication-site hypothesis and its implications for pathogenesis. In Slow Transmissible Diseases of the Nervous System vol 2: pp 13–31 Edited by Prusiner S. B., Hadlow W. J. New York: Academic Press;
    [Google Scholar]
  12. Dickinson A. G., Meikle V. M. H., Fraser H. 1969; Genetical control of the concentration of ME7 scrapie agent in the brain of mice. Journal of Comparative Pathology 79:15–22
    [Google Scholar]
  13. Field E. J., Windsor G. D. 1965; Culture characteristics of scrapie mouse brain. Research in Veterinary Science 6:130–132
    [Google Scholar]
  14. Field E. J., Joyce G., Keith A. 1971; Viral properties of scrapie. Nature New Biology 230:56–57
    [Google Scholar]
  15. Fraser H. 1982; Neuronal spread of scrapie agent and targeting of lesions within the retinotectal pathway. Nature, London 295:149–150
    [Google Scholar]
  16. Gajdusek D. C. 1977; Unconventional viruses and the origin and disappearance of kuru. Science 197:943–960
    [Google Scholar]
  17. Gibson P. E., Bell T. M., Field E. J. 1972; Failure of the scrapie agent to replicate in L5178Y mouse leukaemic cells. Research in Veterinary Science 13:95–96
    [Google Scholar]
  18. Greene L. A., Rein G. 1977a; Release, storage, and uptake of catecholamines by a clonal line of nerve growth factor (NGF) responsive pheochromocytoma cells. Brain Research 129:247–263
    [Google Scholar]
  19. Greene L. A., Rein G. 1977b; Release of [3H]norepinephrine from a clonal line of pheochromocytoma cells (PC12) by nicotinic cholinergic stimulation. Brain Research 138:521–528
    [Google Scholar]
  20. Greene L. A., Rein G. 1977c; Synthesis, storage and release of acetylcholine by a noradrenergic pheochromocytoma cell line. Nature, London 268:349–351
    [Google Scholar]
  21. Greene L. A., Tischler A. S. 1976; Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proceedings of the National Academy of Sciences, U. S. A 73:2424–2428
    [Google Scholar]
  22. Gustafson D. P., Kanitz C. L. 1965; Evidence of the presence of scrapie in cell culture of brain. In Slow, Latent and Temperate Virus Infection pp 221–236 Edited by Gajdusek D. C., Gibbs C. J., Alpers M. National Institute of Neurological Diseases and Blindness, Monograph No. 2. Bethesda, Md.: National Institutes of Health;
    [Google Scholar]
  23. Haig D. A., Clarke M. C. 1965; Observations on the agent of scrapie. In Slow, Latent and Temperate Virus Infections pp 215–219 Edited by Gajdusek D. C., Gibbs C. J., Alpers M. National Institute of Neurological Diseases and Blindness, Monograph No. 2 Bethesda, Md: National Institutes of Health;
    [Google Scholar]
  24. Haig D. A., Pattison I. H. 1967; In vitro growth of pieces of brain from scrapie-affected mice. Journal of Pathology and Bacteriology 93:724–727
    [Google Scholar]
  25. Hunter G. D., Millson G. C., Chandler R. L. 1963; Observations on the comparative infectivity of cellular fractions derived from homogenates of mouse-scrapie brain. Research in Veterinary Science 4:543–549
    [Google Scholar]
  26. Hunter G. D., Kimberlin R. H., Millson G. C. 1972; Absence of eclipse phase in scrapie mice. Nature New Biology 235:31–32
    [Google Scholar]
  27. Kimberlin R. H. 1976; Scrapie in the Mouse. Patterns of Progress, Microbiology Series vol 5: Shildon, Co. Durham: Meadowfield Press;
    [Google Scholar]
  28. Kimberlin R. H. 1979; Early events in the pathogenesis of scrapie in mice: biological and biochemical studies. In Slow Transmissible Diseases of the Nervous System vol 2: pp 33–54 Edited by Prusiner S. B., Hadlow W. J. New York: Academic Press;
    [Google Scholar]
  29. Kimberlin R. H., Walker C. A. 1980; Pathogenesis of mouse scrapie: evidence for neural spread of infection to the CNS. Journal of General Virology 51:183–187
    [Google Scholar]
  30. Kimberlin R. H., Walker C. A. 1982; Pathogenesis of mouse scapie. Patterns of agent replication in different parts of the CNS following intraperitoneal infection. Journal of the Royal Society of Medicine 75:618–624
    [Google Scholar]
  31. McNemar Q. 1955 Psychological Statistics pp 19–20 357–358 New York: John Wiley & Sons;
    [Google Scholar]
  32. Markovits P., Dormont D., Delpeck B., Court L., Latarjet R. 1981; Essais de propagation in vitro de l’agent scrapie dans des cellules nerveuses de souris. Comptes rendus hebdomadaires des séances de l’Académie des sciences 293:413–417
    [Google Scholar]
  33. Markovits P., Dormont D., Maunoury R., Delamarche C., Delpech A., Dianoux L., Latarjet R. 1982; Modifications in vitro de la morphologie et de la croissance de cellules infectees par l’agent scrapie. Comptes rendus hebdomadaires des séances de l’Académie des sciences 294:305–312
    [Google Scholar]
  34. Marsh R. F. 1974; Slow virus diseases of the central nervous system. Advances in Veterinary Science and Comparative Medicine 18:155–178
    [Google Scholar]
  35. Mobley W. C., Schenker A., Shooter E. M. 1976; Characterization and isolation of proteolytically modified nerve growth factor. Biochemistry 15:5543–5552
    [Google Scholar]
  36. Outram G. W. 1976; The pathogenesis of scrapie in mice. In Slow Virus Diseases of Animals and Man, Frontiers of Biology Series vol 44: pp 325–357 Edited by Kimberlin R. H. Amsterdam & Oxford: North-Holland;
    [Google Scholar]
  37. Reiger F., Helanski M. L., Greene L. A. 1980; The effects of nerve growth factor on acetylcholinesterase and its multiple forms in cultures of rat PC12 pheochromocytoma cells: increased total specific activity and appearance of the 16S molecular form. Developmental Biology 76:238–243
    [Google Scholar]
  38. Rubenstein R., Price R. W. 1983a; Preservation of catecholamine uptake and release in herpes simplex virus type 1-infected PC12 cells. Journal of General Virology 64:2505–2509
    [Google Scholar]
  39. Rubenstein R., Price R. W. 1983b; Replication of thymidine kinase deficient herpes simplex virus type 1 in neuronal cell culture: infection of the PC12 cell. Archives of Virology 78:49–64
    [Google Scholar]
  40. Rubenstein R., Price R. W. 1983c; Early inhibition of acetylcholinesterase and choline acetyltransferase activity in herpes simplex virus type 1 infection of PC12 cells. Journal of Neurochemistry 42:142–150
    [Google Scholar]
  41. Rudy B., Kirschenbaum B., Greene L. A. 1982; Nerve growth factor induced increase in saxitoxin binding to rat PC12 pheochromocytoma cells. Journal of Neuroscience 2:1405–1411
    [Google Scholar]
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