Abstract
SUMMARY
1. To elucidate mechanisms for the generation of the detergent-insoluble, proteinase K-resistant prion protein (PrPSc) from the detergent-soluble, proteinase K-sensitive PrP (PrPC) and the replication of the infectious agent in prion diseases, we followed the kinetics of detergent-insoluble PrP and PrPSc levels, infectious titers, and associated pathological changes in the brains of mice inoculated with a mouse-adapted Creutzfeldt–Jakob disease agent.
2. PrPSc in brain homogenate and detergent-insoluble PrP enriched by two-cycle ultracentrifugation were detected by immunoblotting and their relative amounts were estimated according to a standard curve plotted between the amount of PrP and signal intensity on immunoblotting. The titer of infectivity was determined by the incubation periods of mice inoculated with the unfractionated homogenate on the basis of a standard curve plotted between the titer and incubation period.
3. Detergent-insoluble PrP became detectable 4 weeks postinoculation (p.i.) well before the detection of PrPSc. The low level of detergent-insoluble PrP continued until dramatic accumulation occurred at 14 weeks p.i., correlating well with the accumulation of PrPSc and development of pathological changes. The infectious titer was undetectable at 4 weeks p.i. and its logarithmic increase occurred 10 weeks p.i. preceding the logarithmic accumulation of PrPs.
4. The lag time of detergent-insoluble PrP accumulation and the discrepancy between infectious titers and PrPs observed during the early period after inoculation suggest a slow and rate-limiting step for the detergent-insoluble PrP to become the infectious agent-associated PrPSc.
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REFERENCES
Alper, T., Haig, D. A., and Clarke, M. C. (1966). The exceptionally small size of the scrapie agent. Biochem. Biophys. Res. Commun. 22:278-284.
Bruce, M. E., and Fraser, H. (1991). Scrapie strain variation and its implications. Curr. Top. Microbiol. Immunol. 172:125-138.
Büeler, H., Aguzzi, A., Sailer, A., Greiner, R. A., Autenried, P., Aguet, M., and Weissmann, C. (1993). Mice devoid of PrP are resistant to scrapie. Cell 73:1339-1347.
Cohen, F. E., Pan, K. M., Huang, Z., Baldwin, M., Fletterick, R. J., and Prusiner, S. B. (1994). Structural clues to prion replication. Science 264:530-531.
DeArmond, S. J., and Prusiner, S. B. (1993). The neurochemistry of prion diseases. J. Neurochem. 61:1589-1601.
Hill, A. F., Antoniou, M., and Collinge, J. (1999). Protease-resistant prion protein produced in vitro lacks detectable infectivity. J. Gen. Virol. 80:11-14.
Hsiao, K. K., Groth, D., Scott, M., Yang, S. L., Serban, H., Rapp, D., Foster, D., Torchia, M., Dearmond, S. J., and Prusiner, S. B. (1994). Serial transmission in rodents of neurodegeneration from transgenic mice expressing mutant prion protein. Proc. Natl. Acad. Sci. USA 91:9126-9130.
Hsiao, K. K., Scott, M., Foster, D., Groth, D. F., DeArmond, S. J., and Prusiner, S. B. (1990). Spontaneous neurodegeneration in transgenic mice with mutant prion protein. Science 250:1587-1590.
Hunter, G. D. (1969). The size and intracellular location of the scrapie agent. Biochem. J. 114:22-23.
Jarrett, J. P., and Lansbury, P. T., Jr. (1993). Seeding “one-dimensional crystallization” of amyloid: A pathogenic mechanism in Alzheimer's disease and scrapie? Cell 73:1055-1058.
Kaneko, K., Ball, H. L., Wille, H., Zhang, H., Groth, D., Torchia, M., Tremblay, P., Safar, J., Prusiner, S. B., DeArmond, S. J., Baldwin, M. A., and Cohen, F. E. (2000). A synthetic peptide initiates Gerstmann-Sträussler-Scheinker (GSS) disease in transgenic mice. J. Mol. Biol. 295:997-1007.
Kocisko, D. A., Come, J. H., Priola, S. A., Chesebro, B., Raymond, G. J., Lansbury, P. T., and Caughey, B. (1994). Cell-free formation of protease-resistant prion protein. Nature 370:471-474.
Kopacek, J., Sakaguchi, S., Shigematsu, K., Nishida, N., Atarashi, R., Nakaoke, R., Moriuchi, R., Niwa, M., and Katamine, S. (2000). Upregulation of the genes encoding lysosomal hydrolases, a perforin-like protein, and peroxidases in the brains of mice affected with an experimental prion disease. J. Virol. 74:411-417.
Lehmann, S., and Harris, D. A. (1997). Blockade of glycosylation promotes acquisition of scrapie-like properties by the prion protein in cultured cells. J. Biol. Chem. 272:21479-21487.
Manson, J. C., Clarke, A. R., McBride, P. A., McConnell, I., and Hope, J. (1994). PrP gene dosage determines the timing but not the final intensity or distribution of lesions in scrapie pathology. Neurodegeneration 3:331-340.
McKinley, M. P., Bolton, D. C., and Prusiner, S. B. (1983). A protease-resistant protein is a structural component of the scrapie prion. Cell 35:57-62.
Nishida, N., Tremblay, P., Sugimoto, T., Shigematsu, K., Shirabe, S., Petromilli, C., Erpel, S. P., Nakaoke, R., Atarashi, R., Houtani, T., Torchia, M., Sakaguchi, S., DeArmond, S. J., Prusiner, S. B., and Katamine, S. (1999). A mouse prion protein transgene rescues mice deficient for the prion protein gene from Purkinje cell degeneration and demyelination. Lab. Invest. 79:689-697.
Pan, K. M., Baldwin, M., Nguyen, J., Gasset, M., Serban, A., Groth, D., Mehlhorn, I., Huang, Z., Fletterick, R. J., Cohen, F. E., et al. (1993). Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc. Natl. Acad. Sci. USA 90:10962-10966.
Prusiner, S. B. (1982). Novel proteinaceous infectious particles cause scrapie. Science 216:136-144.
Prusiner, S. B. (1991). Molecular biology of prion diseases. Science 252:1515-1522.
Prusiner, S. B. (1994). Biology and genetics of prion diseases. Annu. Rev. Microbiol. 48:655-686.
Prusiner, S. B., Groth, D., Serban, A., Koehler, R., Foster, D., Torchia, M., Burton, D., Yang, S. L., and DeArmond, S. J. (1993). Ablation of the prion protein (PrP) gene in mice prevents scrapie and facilitates production of anti-PrP antibodies. Proc. Natl. Acad. Sci. USA 90:10608-10612.
Prusiner, S. B., Scott, M. R., DeArmond, S. J., and Cohen, F. E. (1998). Prion protein biology. Cell 93:337-348.
Sakaguchi, S., Katamine, S., Shigematsu, K., Nakatani, A., Moriuchi, R., Nishida, N., Kurokawa, K., Nakaoke, R., Sato, H., Jishage, K., Kuno, J., Noda, T., and Miyamoto, T. (1995). Accumulation of proteinase K-resistant prion protein (PrP) is restricted by the expression level of normal PrP in mice inoculated with a mouse-adapted strain of the Creutzfeldt-Jakob disease agent. J. Virol. 69:7586-7592.
Sakaguchi, S., Katamine, S., Yamanouchi, K., Kishikawa, M., Moriuchi, R., Yasukawa, N., Doi, T., and Miyamoto, T. (1993). Kinetics of infectivity are dissociated from PrP accumulation in salivary glands of Creutzfeldt-Jakob disease agent-inoculated mice. J. Gen. Virol. 74:2117-2123.
Tateishi, J., Ohta, M., Koga, M., Sato, Y., and Kuroiwa, Y. (1979). Transmission of chronic spongiform encephalopathy with kuru plaques from humans to small rodents. Ann. Neurol. 5:581-584.
Telling, G. C., Scott, M., Mastrianni, J., Gabizon, R., Torchia, M., Cohen, F. E., DeArmond, S. J., and Prusiner, S. B. (1995). Prion propagation in mice expressing human and chimeric PrP transgenes implicates the interaction of cellular PrP with another protein. Cell 83:79-90.
Weissmann, C. (1991). A 'unified theory' of prion propagation. Nature 352:679-683.
Xi, Y. G., Ingrosso, L., Ladogana, A., Masullo, C., and Pocchiari, M. (1992). Amphotericin B treatment dissociates in vivo replication of the scrapie agent from PrP accumulation. Nature 356:598-601.
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Nakaoke, R., Sakaguchi, S., Atarashi, R. et al. Early Appearance but Lagged Accumulation of Detergent-Insoluble Prion Protein in the Brains of Mice Inoculated with a Mouse-Adapted Creutzfeldt–Jakob Disease Agent. Cell Mol Neurobiol 20, 717–730 (2000). https://doi.org/10.1023/A:1007054909662
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DOI: https://doi.org/10.1023/A:1007054909662