Dottore di ricerca

ciclo: XXXII

Titolo della tesi: The neurotoxicity of native metastable prefibrillar oligomers from salmon Calcitonin: an innovative amyloid-induced excitotoxicity paradigm

Protein misfolding is implicated in different severe amyloid related neurodegenerative diseases, like Alzheimer, Parkinson and Creutzfeldt-Jacob. This process results in the aggregation of toxic small soluble prefibrillar oligomers (PFOs), almost ineffective proto-fibrils (PFs) and mature fibres (MFs). Many efforts are pointing to clarify PFOs structures and cytotoxicity. However, since their metastability, oligomers change their conformation along the experiments leading to inconsistency in comparing data from different groups. Salmon Calcitonin (sCT), is a 32 amino acid polypeptide hormone displaying the tendency to self-assemble in the amyloid pattern, with a very low aggregation rate. This appealing feature allows to investigate the effects of native PFOs in the early stages of neurotoxicity, without any chemical stabilization. We purified sCT PFOs enriched fractions by means of size exclusion chromatography (SEC) aiming to find who is the most toxic species, among sCT PFOs, comparing the biologic effects of native PFOs enriched fraction with respect to sCT monomers. The intracellular Ca2+ rise plays a fundamental role in amyloid protein-induced neurodegenerations. According to what we found in literature, two paradigms have been explored: i) the “membrane permeabilization” due to the formation of amyloid pores or other types of membrane damage; ii) “receptor-mediated” modulation of Ca2+ channels. Therefore, we tested the effects of native sCT PFOs or monomers in differentiated neurons, by means of Ca2+imaging, cellular viability, Long-Term Potentiation, Post-Synaptic Densities protein expression, excised patch clamp recordings and miniature-EPSC. Results indicated that PFOs-, but not Monomer-enriched solutions, induced abnormal permeability to Ca2+, not fully explained by NMDARs activation. We also found that, the formation of small amyloid pores was consistent with an increased quantal release of neurotransmitters, able to drive a receptor mediated response, per se. Thus, we propose an innovative neurotoxicity mechanism for amyloid proteins where the “membrane permeabilization” and “receptor-mediated” paradigms coexist.

Produzione scientifica

11573/1290866 - 2019 - Calcitonin native prefibrillar oligomers but not monomers induce membrane damage that triggers NMDA-mediated Ca 2+ -influx, LTP impairment and neurotoxicity
Belfiore, Marcello; Cariati, I.; Matteucci, A.; Gaddini, L.; Macchia, G.; Fioravanti, R.; Frank, C.; Tancredi, V.; D'arcangelo, G.; Diociaiuti, M. - 01a Articolo in rivista
rivista: SCIENTIFIC REPORTS (London: Springer Nature London: Nature Publishing Group) pp. - - issn: 2045-2322 - wos: WOS:000462298600031 (10) - scopus: 2-s2.0-85063522872 (12)

11573/1345440 - 2019 - The Interaction between Amyloid Prefibrillar Oligomers of Salmon Calcitonin and a Lipid-Raft Model: Molecular Mechanisms Leading to Membrane Damage, Ca2+-Influx and Neurotoxicity
Diociaiuti, Marco; Bombelli, Cecilia; Zanetti-Polzi, Laura; Belfiore, Marcello; Fioravanti, Raoul; Macchia, Gianfranco; Giordani, Cristiano - 01a Articolo in rivista
rivista: BIOMOLECULES (Basel: MDPI) pp. - - issn: 2218-273X - wos: WOS:000514863200123 (3) - scopus: 2-s2.0-85077551718 (3)

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