Abstract
Prion neurotoxicity has been modeled in vitro using synthetic peptides derived from the PrPC sequence. The major region of neurotoxicity has been localized to the hydrophobic domain located in the middle of the PrP protein. Neurotoxicity assays are typically performed on cultured mouse cerebellar neurons derived from neonatal pups, and cell viability can be monitored by assays including MTT or MTS, cell death by LDH release, or apoptosis by caspase cleavage assays. These neurotoxicity studies have been useful in identifying cofactors, such as PrPC and metals, as modulators of PrP peptide-mediated neurotoxicity. Given the biosafety issues associated with handling and purifying infectious prions, the use of synthetic peptides, which display a dependence upon PrPC expression for toxicity, as per the PrPSc agent for infectivity, supports the relevance of using these synthetic peptides for understanding PrP-mediated neurotoxicity.
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References
Tagliavini F, Prelli F, Verga L et al (1993) Synthetic peptides homologous to prion protein residues 106-147 form amyloid-like fibrils in vitro. Proc Natl Acad Sci U S A 90:9678–9682
Forloni G, Angeretti N, Chiesa R et al (1993) Neurotoxicity of a prion protein fragment. Nature 362:543–546
Thompson A, White AR, McLean C et al (2000) Amyloidogenicity and neurotoxicity of peptides corresponding to the helical regions of PrP(C). J Neurosci Res 62:293–301
Brown DR (2000) Prion protein peptides: optimal toxicity and peptide blockade of toxicity. Mol Cell Neurosci 15:66–78
Bergstrom AL, Cordes H, Zsurger N et al (2005) Amidation and structure relaxation abolish the neurotoxicity of the prion peptide PrP106-126 in vivo and in vitro. J Biol Chem 280:23114–23121
Thellung S, Florio T, Corsaro A et al (2000) Intracellular mechanisms mediating the neuronal death and astrogliosis induced by the prion protein fragment 106-126. Int J Dev Neurosci 18:481–492
White AR, Maher F, Brazier MW et al (2003) Diverse fibrillar peptides directly bind the Alzheimer's amyloid precursor protein and amyloid precursor-like protein 2 resulting in cellular accumulation. Brain Res 966:231–244
White AR, Guirguis R, Brazier MW et al (2001) Sublethal concentrations of prion peptide PrP106-126 or the amyloid beta peptide of Alzheimer's disease activates expression of proapoptotic markers in primary cortical neurons. Neurobiol Dis 8:299–316
Forloni G, Bugiani O, Tagliavini F, Salmona M (1996) Apoptosis-mediated neurotoxicity induced by beta-amyloid and PrP fragments. Mol Chem Neuropathol 28:163–171
Brown DR, Herms J, Kretzschmar HA (1994) Mouse cortical cells lacking cellular PrP survive in culture with a neurotoxic PrP fragment. Neuroreport 5:2057–2060
White AR, Collins SJ, Maher F et al (1999) Prion protein-deficient neurons reveal lower glutathione reductase activity and increased susceptibility to hydrogen peroxide toxicity. Am J Pathol 155:1723–1730
Jobling MF, Huang X, Stewart LR et al (2001) Copper and zinc binding modulates the aggregation and neurotoxic properties of the prion peptide PrP106-126. Biochemistry 40:8073–8084
Stewart LR, White AR, Jobling MF et al (2001) Involvement of the 5-lipoxygenase pathway in the neurotoxicity of the prion peptide PrP106-126. J Neurosci Res 65:565–572
Salmona M, Malesani P, De Gioia L et al (1999) Molecular determinants of the physicochemical properties of a critical prion protein region comprising residues 106-126. Biochem J 342:207–214
Jobling MF, Stewart LR, White AR et al (1999) The hydrophobic core sequence modulates the neurotoxic and secondary structure properties of the prion peptide 106-126. J Neurochem 73:1557–1565
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Ciccotosto, G.D., Jana, M., Cappai, R. (2017). Neurotoxicity of Prion Peptides on Cultured Cerebellar Neurons. In: Lawson, V. (eds) Prions. Methods in Molecular Biology, vol 1658. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7244-9_12
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DOI: https://doi.org/10.1007/978-1-4939-7244-9_12
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