Cellular and Molecular Neurobiology

, Volume 25, Issue 8, pp 1171–1183 | Cite as

Altered Expression of the Prion Gene in Rat Astrocyte and Neuron Cultures Treated with Prion Peptide 106–126

  • Zhang-Yong Ning
  • De-Ming Zhao
  • Hong-Xiang Liu
  • Jian-Min Yang
  • Cai-Xia Han
  • Ya-Li Cui
  • Li-Ping Meng
  • Chang-De Wu
  • Mei-Li Liu
  • Tai-Xiang Zhang
Article
Received:
Accepted:

Summary

Neuronal degeneration and astrogliosis are hallmarks of prion disease. Synthetic prion protein (PrP) peptide 106–126 (PrP106–126) can induce death of neurons and proliferation of astrocytes in vitro and this neurotoxic effect depends on the expression of cellular PrP (PrPC) and is hence believed to be PrPC -mediated. To further elucidate the involvement of PrPC in PrP106–126-induced neurotoxicity, we determined the expression of PrP mRNA in primary culture of rat cortical neuron cells, cerebellar granule cells, and astrocytes following treatment with 50μM of PrP106–126 scrambled PrP106–126 by quantitative real-time RT-PCR. As shown by MTT test, PrP106–126 induced significant death of neuron cells and marked proliferation of astrocytes after 10 days of treatment. Under the same treatment regimens, the level of PrP gene expression was significantly down-regulated in cortical neuron cell cultures and cerebellar granule cell cultures and was up-regulated in astrocyte cultures. The altered PrP gene expression occurred as early as 3 days after the treatment. After 10 days of treatment, while the cultured cortical neurons underwent further apoptosis, their expression of PrP gene started to recover gradually. These findings indicate that PrP 106–126 regulates transcription of the PrP gene and this activity is associated with its neurotoxicity in primary rat neuronal cultures.

Keywords

prio npeptide 106–126 cultured astrocytes and neurons mRNA expression real-time RT-PCR 

Abbreviation

bp

base pairs

BSE

bovine spongiform encephalopathy

cDNA

complementary deoxyribonucleic acid

CJD

Creutzfeldt–Jakob disease

GPI

glycoinositol phospholipid

GSS

Gerstmann–Sträussler–Scheinker disease

mRNA

messenger ribonucleic acid

PCR

polymerase chain reaction

PrP

prion protein

PrPC

cellular PrP

PrPSC

scrapie isoform of PrP

RT–PCR

reverse transcription polymerase chain reaction

TSE

transmissible spongiform encephalopathy.

This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Blattler, T., Brandner, S., Raeber, A. J., Klein, M. A., Voigtlander, T., Weissmann, C., and Aguzzi, A. (1997). PrP-expressing tissue required for transfer of scrapie infectivity from spleen to brain. Nature 389:69–73.PubMedGoogle Scholar
  2. Brown, D. R. (1999a). Prion protein peptide neurotoxicity can be mediated by astrocytes. J. Neurochem. 73(3):1105–1113.CrossRefGoogle Scholar
  3. Brown, D. R. (1999b). Neurons depend on astrocytes in a coculture system for protection from glutamate toxicity. Mol. Cell. Neurosci. 13(5):379–389.CrossRefGoogle Scholar
  4. Brown, D. R. (2000). PrPSc-like prion protein peptide inhibits the function of cellular prion protein. Biochem. J. 352:511–518.CrossRefPubMedGoogle Scholar
  5. Brown, D. R. (2001). Prion and prejudice: Normal protein and the synapse. Trends Neurosci. 24(2):85–90.CrossRefPubMedGoogle Scholar
  6. Brown, D. R., and Mohn, C. M. (1999). Astrocytic glutamate uptake and prion protein expression. Glia 25(3):282–292.CrossRefPubMedGoogle Scholar
  7. Brown, D. R., Schmidt, B., and Kretzschmar, H. A. (1996). Role of microglia and host prion protein in neurotoxicity of a prion protein fragment. Nature 380:345–347.PubMedGoogle Scholar
  8. Bruce, M. E., McBride, P. A., and Farquhar, C. F. (1989). Precise targeting of the sialoglyco-protein PrP and vacuolar degeneration in mouse scrapie. Neurosci. Lett. 102(1):1–6.CrossRefPubMedGoogle Scholar
  9. Chabry, J., Ratsimanohatra, C., Sponne, I., Elena, P. P., Vincent, J. P., and Pillot, T. (2003). In vivo and in vitro neurotoxicity of the human prion protein (PrP) fragment P118–135 independently of PrP expression. J. Neurosci. 23:462–469.PubMedGoogle Scholar
  10. Chen, S. G., Teplow, D. B., Parchi, P., Teller, J. K., Gambetti, P., and Autilio-Gambetti, L. (1995). Truncated forms of the human prion protein in normal brain and in prion diseases. J. Biol. Chem. 270(32):19173–19180.PubMedGoogle Scholar
  11. Chiesa, R., and Harris, D. A. (2001). Prion diseases: What is the neurotoxic molecule? Neurobiol. Dis. 8(5):743–763.CrossRefPubMedGoogle Scholar
  12. Collinge, J. (2001). Prion diseases of humans and animals: Their causes and molecular basis. Annu. Rev. Neurosci. 24:519–550.CrossRefPubMedGoogle Scholar
  13. DeArmond, S. J., Mobley, W. C., DeMott, D. L., Barry, R. A., Beckstead, J. H., and Prusiner, S. B. (1987). Changes in the localization of brain prion proteins during scrapie infection. Neurology 37(8):1271–1280.PubMedGoogle Scholar
  14. Diedrich, J. F., Bendheim, P. E., Kim, Y. S., Carp, R. I., and Haase, A. T. (1991). Scrapie-associated prion protein accumulates in astrocytes during scrapie infection. Proc. Natl. Acad. Sci. U.S.A. 88(2):375–379.PubMedGoogle Scholar
  15. Ettaiche, M., Pichot, R., Vincent, J. P., and Chabry, J. (2000). In vivo cytotoxicity of the prion protein fragment 106–126. J. Biol. Chem. 275(47):36487–36490.CrossRefPubMedGoogle Scholar
  16. Fioriti, L., Quagliom, E., Massignan, T., Colombo, L., Stewart, R. S., Salmona, M., Harris, D. A., Forloni, G., and Chiesa, R. (2005). The neurotoxicity of prion protein (PrP) peptide 106–126 is independent of the expression level of PrP and is not mediated by abnormal PrP species. Mol. Cell. Neurosci. 28(1):165–176.CrossRefPubMedGoogle Scholar
  17. Florio, T., Thellung, S., Amico, C., Robello, M., Salmona, M., Bugiani, O., Tagliavini, F., Forloni, G., and Schettini, G. (1998). Prion protein fragment 106–126 induces apoptotic cell death and impairment of L-type voltage-sensitive calcium channel activity in the GH3 cell line. J. Neurosci. Res. 54(3):341–352.CrossRefPubMedGoogle Scholar
  18. Forloni, G., Angeretti, N., Chiesa, R., Monzani, E., Salmona, M., Bugiani, O., and Tagliavini, F. (1993). Neurotoxicity of a prion protein fragment. Nature 362:543–546.CrossRefPubMedGoogle Scholar
  19. Forloni, G., Del, B. R., Angeretti, N., Chiesa, R., Smiroldo, S., Doni, R., Ghibaudi, E., Salmona, M., Porro, M., Verga, L., Giaccone, G., Bugiani, O., and Tagliavini, F. (1994). A neurotoxic prion protein fragment induces rat astroglial proliferation and hypertrophy. Eur. J. Neurosci. 6(9):1415–1422.PubMedGoogle Scholar
  20. Freshney, R. I. (1987). Culture of specific cell types. In Freshney, R. I. (ed.), Culture of Animal Cells: A Manual of Basic Technique, AR Liss, New York, pp. 257–288.Google Scholar
  21. Glatzel, M., and Aguzzi, A. (2000). PrP(C) expression in the peripheral nervous system is a determinant of prion neuroinvasion. J. Gen. Virol. 81:2813–2821.PubMedGoogle Scholar
  22. Gu, Y., Fujioka, H., Mishra, R. S., Li, R., and Singh, N. (2002). Prion peptide 106–126 modulates the aggregation of cellular prion protein and induces the synthesis of potentially neurotoxic transmembrane PrP. J. Biol. Chem. 277:2275–2286.PubMedGoogle Scholar
  23. Hope, J., Shearman, M. S., Baxter, H. C., Chong, A., Kelly, S. M., and Price, N. C. (1996). Cytotoxicity of prion protein peptide (PrP106–126) differs in mechanism from the cytotoxic activity of the Alzheimer's disease amyloid peptide, a beta 25–35. Neurodegeneration 5(1):1–11.CrossRefPubMedGoogle Scholar
  24. Levi, G., Aloisi, F., Ciotti, M. T., and Gallo, V. (1984). Autoradiographic localization and depolarization-induced release of acidic amino acids in differentiating cerebellar granule cell cultures. Brain Res. 290(1):77–86.CrossRefPubMedGoogle Scholar
  25. Liemann, S., and Glockshuber, R. (1998). Transmissible spongiform encephalopathies. Biochem. Biophys. Res. Commun. 250(2):187–193.CrossRefPubMedGoogle Scholar
  26. Prusiner, S. B. (1991). Molecular biology of prion diseases. Science 252:1515–1522.PubMedGoogle Scholar
  27. Prusiner, S. B. (1992). Chemistry and biology of prions. Biochemistry 31(49):12277–12288.CrossRefPubMedGoogle Scholar
  28. Prusiner, S. B. (1998). Prions. Proc. Natl. Acad. Sci. U.S.A. 95(23):13363–13383.CrossRefPubMedGoogle Scholar
  29. Raeber, A. J., Race, R. E., Brandner, S., Priola, S. A., Sailer, A., Bessen, R. A., Mucke, L., Manson, J., Aguzzi, A., Oldstone, M. B., Weissmann, C., and Chesebro, B. (1997). Astrocyte-specific expression of hamster prion protein (PrP) renders PrP knockout mice susceptible to hamster scrapie. EMBO J. 16(20):6057–6065.CrossRefPubMedGoogle Scholar
  30. Rymer, D. L., and Good, T. A. (2000). The role of prion peptide structure and aggregation in toxicity and membrane binding. J. Neurochem. 75(6):2536–2545.CrossRefPubMedGoogle Scholar
  31. Selvaggini, C., De Gioia, L., Cantu, L., Ghibaudi, E., Diomede, L., Passerini, F., Forloni, G., Bugiani, O., Tagliavini, F., and Salmona, M. (1993). Molecular characteristics of a protease-resistant, amyloidogenic and neurotoxic peptide homologous to residues 106–126 of the prion protein. Biochem. Biophys. Res. Commun. 194(3):1380–1386.CrossRefPubMedGoogle Scholar
  32. Thellung, S., Florio, T., Villa, V., Corsaro, A., Arena, S., Amico, C., Robello, M., Salmona, M., Forloni, G., Bugiani, O., Tagliavini, F., and Schettini, G. (2000). Apoptotic cell death and impairment of L-type voltage-sensitive calcium channel activity in rat cerebellar granule cells treated with the prion protein fragment 106–126. Neurobiol. Dis. 7(4):299–309.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Zhang-Yong Ning
    • 1
  • De-Ming Zhao
    • 1
    • 3
  • Hong-Xiang Liu
    • 2
  • Jian-Min Yang
    • 1
  • Cai-Xia Han
    • 1
  • Ya-Li Cui
    • 1
  • Li-Ping Meng
    • 1
  • Chang-De Wu
    • 1
  • Mei-Li Liu
    • 1
  • Tai-Xiang Zhang
    • 1
  1. 1.National Animal Transmissible Spongiform Encephalopathies LaboratoryCollege of Veterinary Medicine, China Agricultural UniversityBeijingPeople's Republic of China
  2. 2.Department of PathologyUniversity of CambridgeCambridgeUnited Kingdom
  3. 3.National Animal Transmissible Spongiform Encephalopathies LaboratoryCollege of Veterinary Medicine, China Agricultural UniversityBeijingPeople's Republic of China

Personalised recommendations