Abstract
The neurotoxic activity of β-amyloid (βA) and prion protein (PrP) fragments contributed to the hypothesis concerning a causal role of amyloid deposits in Alzheimer disease (AD) and in prion-related encephalopathies. In this study, we investigated some aspects of the molecular mechanisms associated with neurotoxic activity of synthetic peptides homologous to βA (β 25–35) or PrP (PrP106–126) fragments. Chronic (5–7 d) exposure to both peptides induced neuronal death by apoptosis, as suggested by biochemical and morphological analysis. The apoptotic mechanism was confirmed by ultrastructural examination. The intracellular cascade of events activated by peptides was investigated by Northern blot and PCR analysis of expression of early genes (c-fos, c-jun, c-myc) and other proteins (p53, SGP-2bcl-2, HSP70, Ich-1) potentially involved in apoptosis. With the exception of bcl-2 mRNA decrease and a slight increase of SGP-2 in PrP106–126-treated cells, no consistent alterations of these mRNA expressions were found in neuronal cells exposed to β 25–35 or PrP106–126. Furthermore, we synthesized amidated homologs of both peptides with low amyloidogenic activity to test directly the relationship between amyloid fibrils and cell death. The neurotoxicity exhibited by PrP106–126-NH2 was similar to that observed with original peptide, whereas the amidation of β 25–35 partially reduced the neurotoxicity of this peptide.
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Allsopp T. E., Xyatt S., Paterson H. F., and Davies A. M. (1993) The protooncogene bcl-2 can selectively rescue neurotrophic factor dependent neurons from apoptosis.Cell 73, 295–307.
Chiesa R., Angeretti N., Lucca, Salmona M., Tagliavini F., Bugiani O., and Forloni G. (1996) Clusterin (SGP-2) induction in rat astroglial cells exposed to prion protein fragment 106–126.Eur. J. Neurosci. 8, 589–597.
Chomczynsky P. and Sacchi W. (1987) Single-step method of RNA isolation by acid guandidium thiocyanate-phenol-chloroform extraction.Anal. Biochem. 162, 156–159.
De Armond S. J. and Prusiner S. B. (1995) Etiology and pathogenesis of prion diseases.Am. J. Pathol. 146, 785–811.
De Armond S. J., Mobley W. C., DeMott D. L., Barry R. A., and Beckstead J. H. (1987) Changes in the localization of brain prion proteins during scrapie infection.Neurology 37, 1271–1280.
Duguid J. R., Bohmont C. W., Liu N., and Tourtellotte W. W. (1989) Changes in brain gene expression shared by scrapie and Alzheimer, disease.Proc. Natl. Acad. Sci. USA 86, 7260–7264.
Forloni G., Demicheli F., Giorgi S., Bendotti C., and Angeretti N. (1992) Expression of Amyloid precursor protein mRNAs in endothelial, neuronal and glial cells: modulation by interleukin-1.Mol. Brain Res. 16, 128–134.
Forloni G., Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O., and Tagliavini F. (1993a) Neurotoxicity of a prion protein fragment.Nature 362, 543–546.
Forloni G., Chiesa R., Smiroldo S., Verga L., Salmona M., Tagliavini F., and Angeretti N. (1993b) Apoptosis mediated neurotoxicity induced by chronic application of beta amyloid fragment 25–35.NeuroReport 9, 523–526.
Forloni G. (1993) Neurotoxicity of β amyloid.Funct. Neurol. 8, 211–225.
Forloni G., Del Bo 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 astroglial proliferation and hypertrophy.Eur. J. Neurosci. 6, 1415–1422.
Ghetti B., Tagliavini F., Masters C. L., Beyreuther K., Giaccone G., Verga L., Farlow M. R., Conneally P. M., Dlouly S. R., Azzarelli B., and Bugiani O. (1989) Gerstmann-Sträussler-Scheinker disease. II Neurofibrillary tangles and plaques with PrP-amyloid coexist in an affected family.Neurology 39, 1453–1461.
Goate, Chartier-Harlin M. C., Mullam M., Brown, J., Crawford F., Fidani L., Giuffra L., Haynes A., Irving N., James L., Mant R., Newton P., Rooke K., Roques P., Talbot C., Pericak-Vance M., Roses A., Williamson R., Rossor M., Owen M., and Hardy J. (1991) Segregation of missense mutation in the amyloid precursor protein gene with familial Alzheimer's disease.Nature 349, 704–706.
Loo D. T., Copani A., Pike C. J., Whittmore E. R., Walencewicz A., and Cotman C. W. (1993) Apoptosis is induced by β-amyloid in cultured CNS neurons.Proc. Natl. Acad. Sci. USA 90, 7951–7955.
Martin D. P., Ito A., Horigome K., Lampe P., and Johnson E. M., Jr. (1992) Biochemical characterization of programmed cell death in NGF-deprived sympathetic neurons.J. Neurobiol. 23, 1205–1220.
Masseroli M., Messori A., Ponti M., Bendotti C., and Forloni G. (1993) Automatic quantitative evaluation of autoradiographic band films by computerized image analysis.Life Sci. 53, 331–336.
Pike C. J., Walencewicz A. J., Glabe C. G., and Cotman C. W. (1991) Aggregation-related toxicity of synthetic β-amyloid protein in hippocampal cultures.Eur. J. Pharmacol. 207, 367–368.
Pike C. J., Burdick D., Walencewicz A. J., Glabe C. G., and Cotman C. W. (1993) Neurodegeneration induced by β-amyloid peptides in vitro: the role of peptide assembly state.J. Neurosci. 13, 1676–1687.
Selkoe D. J. (1991) The molecular pathology of Alzheimer's disease.Neuron 6, 487–498.
Tagliavini F., Prelli F., Ghiso J., Bugiani O., Serban D., Prusiner S. B., Farlow M., Ghetti B., and Frangione B. (1991) Amyloid protein of Gerstmann-Straussler-Scheinker disease (Indiana Kindred) is an 11 kd fragment of prion protein with an N-terminal glycine at codon 58.EMBO J. 10, 513–519.
Tagliavini F., Prelli F., Verga L., Giaccone G., Sarma R., Gorevic P., Ghetti B., Passerini F., Ghibaudi E., Forloni G., Salmona M., Bugiani O., and Frangione B. (1993) Synthetic peptides homologous to prion protein residues 106–147 form amyloid-like fibrils in vitro.Proc. Natl. Acad. Sci. USA 90, 9678–9682.
Tagliavini F., Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O., and Forloni G. (1994) A prion protein peptides induces neuronal death by apoptosis, inAmyloid and Amyloidosis 1993 (Kiselevsky, R., Benson M. D., Frangione B., Gauldier J., Muckle T. J., and Young I. D., eds.), Parthenon, New York, pp. 368–370.
Terzi E., Hólzemann G., and Seelig J. (1994) Alzheimer β-amyloid peptide 25–35: Electrostatic interactions with phospholipid membranes.Biochemistry 33, 7434–7441.
Tomei L. D. (1991) Apoptosis: A program for death or survival? inApoptosis: The Molecular Basis of Cell Death (Tomei L. D. and Cope F. O., eds.), Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY.
Wang L., Miura M., Bergeron L., Zhu H., and Yuan J. (1994) Ich-1 an Ice/ced-relative gene, encodes both positive and negative regulators of programmed cell death.Cell 78, 739–750.
Wyllie A. H., Kerr L. A. M., and Curric A. R. (1980) Cell death: The significance of apoptosis.Inter. Rev. Cytol. 68, 251–307.
Yankner B. A., Duffy L. K., and Kirschner D. A. (1990) Neurotrophic neurotoxic effects of amyloid β protein: reversal by tachynin neuropeptides.Science 250, 279–282.
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Forloni, G., Bugiani, O., Tagliavini, F. et al. Apoptosis-mediated neurotoxicity induced by β-amyloid and PRP fragments. Molecular and Chemical Neuropathology 28, 163–171 (1996). https://doi.org/10.1007/BF02815218
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DOI: https://doi.org/10.1007/BF02815218