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Analysis of autophagy flux during protein aggregation containing alpha-synuclein and hyperphosphorylated tau in cultured cells from hipoccampus, substantia nigra and locus coeruleus

Grant number: 16/14513-8
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): September 01, 2016
Effective date (End): August 31, 2018
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Merari de Fátima Ramires Ferrari
Grantee:Rafaela Regina Cardoso
Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil


Changes in cell structure and biochemistry, formation of senile plaques and neurofibrillary tangles are common features of the brain in degenerative process. These neurofibrillary tangles are consisted essentially of hyperphosphorylated tau protein. The presence of these protein aggregates leads to intracellular damage culminating in cell death. Failure in the process of cellular degradation could favor the aggregation of intracellular and extracellular proteins and culminate in neurodegeneration. For this reason, we intend to analyze the formation of autophagolysosomes and the colocalization of autophagosomes with hyperphosphorylated tau and alpha-synuclein, in order to study the relation between autophagy and protein aggregation. For this analysis, specific antibodies will be used to detect hyperphosphorylated tau and alpha-synuclein. The analysis of autophagolysosomes will be performed using fluorescent probes which mark acidic organelles with different colors depending upon the pH range. This study will be performed on primary cell cultures of the hippocampus, substantia nigra and locus coeruleus subjected to rotenone exposure in concentrations from 0.3 to 1nM, considering that 1nM is able to induce aggregates containing hyperphosphorylated tau protein and alpha-synuclein in the cells.

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
SEMEANO, ANA T.; TOFOLI, FABIANO A.; CORREA-VELLOSO, JULIANA C.; DE JESUS SANTOS, ANA P.; OLIVEIRA-GIACOMELLI, AGATHA; CARDOSO, RAFAELA R.; PESSOA, MATEUS A.; DA ROCHA, EDROALDO LUMMERTZ; RIBEIRO, GUSTAVO; FERRARI, MERARI F. R.; et al. Effects of Magnetite Nanoparticles and Static Magnetic Field on Neural Differentiation of Pluripotent Stem Cells. STEM CELL REVIEWS AND REPORTS, v. 18, n. 4, p. 18-pg., . (18/07366-4, 19/26852-0, 16/14513-8, 13/08028-1, 18/07592-4, 13/08135-2)

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