Effects of miro-1 silencing upon mitophagy and neuronal death during alpha-synucle...
| Grant number: | 16/09621-6 |
| Support type: | Scholarships in Brazil - Scientific Initiation |
| Effective date (Start): | August 01, 2016 |
| Effective date (End): | July 31, 2017 |
| Field of knowledge: | Biological Sciences - Morphology |
| Principal Investigator: | Merari de Fátima Ramires Ferrari |
| Grantee: | Thaís Alexandre Falkembach Andreis |
| Home 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. | |