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Analysis of the role of miro-1 upon alpha-synuclein toxicity in a cellular model of Parkinson's Disease

Grant number: 16/09621-6
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): August 01, 2016
Effective date (End): July 31, 2017
Field of knowledge:Biological Sciences - Morphology - Cytology and Cell Biology
Principal Investigator:Merari de Fátima Ramires Ferrari
Grantee:Thaís Alexandre Falkembach Andreis
Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The presence of non-functional mitochondria in neurons culminates with the signaling for cell death. Mitochondria are recycled with the aid of motor and acessory specific proteins, such as Miro-1. However, apparently Miro-1 is not linked exclusively to the mitochondrial traffic but also relates to the cytoskeleton, endoplasmic reticulum and calcium homeostasis. Recent data from our laboratory indicate that the absence of Miro, in yeast, protects these cells from the toxicity promoted by the expression of alpha-synuclein (a protein associated with Parkinson's disease and other synucleinopathies). The objective of this study is to evaluate the effects of suppression of Miro-1 expression by CRISPR/Cas9 in neurons derived from neuroblastoma, expressing wild-type and mutant (A30P and A53T) alpha-synuclein, specially with respect to cell death, stress endoplasmic reticulum and mitofagia. Therefore, TUNELand MTT assays will be made, levels of activated caspase3 (for evaluation of cell death); CHOP ATF6 and specific probe to the endoplasmic reticulum (for stress and fragmentation analysis of the endoplasmic reticulum); and PINK and parkin (for analysis of mitophagy), thereby allowing study of the mechanism by which Miro-1 can influence the toxicity of alpha-synuclein.

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