Parkinson's disease (PD) is a common and debilitating neurodegenerative disorder, resulting in substantial social and economic hardships. Current therapy is purely symptomatic and loses efficacy over time, as there is no known way to slow the underlying disease progression. Although we do not understand why cells degenerate in PD, multiple lines of evidence indicate that the protein alpha-synuclein and mitochondrial dysfunction play central roles in this process. We have found that alpha-synuclein normally binds to mitochondria and produces a dramatic change in their morphology, suggesting a cellular function and mechanism that might contribute to disease. This proposal will determine the mechanism by which alpha-synuclein binds and induces morphologic changes in mitochondria, as well as the consequences of these changes to cells that degenerate in PD.
In Specific Aim 1, we will use complementary approaches to characterize the effects of alpha-synuclein on mitochondrial morphology. These will include electron microscopy to visualize the ultrastructural effects of alpha-synuclein on mitochondrial morphology, and cellular approaches in which we test whether known inhibitors can block alpha-synuclein from producing morphologic change.
In Specific Aim 2, we will determine the molecular mechanisms by which alpha-synuclein binds to mitochondria and produces morphologic changes. First, we will determine the mitochondrial compartment to which alpha-synuclein localizes. We will then identify and compare the specific domains of alpha-synuclein that are required for binding and morphologic change. We will also assess the effects of known PD mutations, and determine the multimeric state of synuclein that produces the effect.
Specific Aim 3 investigates the consequences of alpha-synuclein's interactions with mitochondria in neurons, by examining whether alpha-synuclein changes the distribution and mobility of mitochondria at the nerve terminal. We will also determine whether interactions with mitochondria contribute to alpha-synuclein's known effects of sensitizing to mitochondrial toxins. The results will contribute to our understanding of PD pathogenesis, and may lead to the development of new diagnostic and therapeutic approaches.

Public Health Relevance

The proposed experiments will provide critical information concerning alpha-synuclein's effects on mitochondria and the consequences of this interaction. The results will provide new insights into why Parkinson's disease occurs, and may form the basis for the development of new diagnostic assays and disease modifying therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08NS062954-05
Application #
8289649
Study Section
NST-2 Subcommittee (NST)
Program Officer
Sutherland, Margaret L
Project Start
2009-07-01
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
5
Fiscal Year
2012
Total Cost
$190,458
Indirect Cost
$14,108
Name
J. David Gladstone Institutes
Department
Type
DUNS #
099992430
City
San Francisco
State
CA
Country
United States
Zip Code
94158
Skibinski, Gaia; Nakamura, Ken; Cookson, Mark R et al. (2014) Mutant LRRK2 toxicity in neurons depends on LRRK2 levels and synuclein but not kinase activity or inclusion bodies. J Neurosci 34:418-33
Berthet, Amandine; Margolis, Elyssa B; Zhang, Jue et al. (2014) Loss of mitochondrial fission depletes axonal mitochondria in midbrain dopamine neurons. J Neurosci 34:14304-17
Itoh, Kie; Nakamura, Ken; Iijima, Miho et al. (2013) Mitochondrial dynamics in neurodegeneration. Trends Cell Biol 23:64-71
Nakamura, Ken (2013) *-Synuclein and mitochondria: partners in crime? Neurotherapeutics 10:391-9
Hertz, Nicholas T; Berthet, Amandine; Sos, Martin L et al. (2013) A neo-substrate that amplifies catalytic activity of parkinson's-disease-related kinase PINK1. Cell 154:737-47
Nakamura, Ken; Nemani, Venu M; Azarbal, Farnaz et al. (2011) Direct membrane association drives mitochondrial fission by the Parkinson disease-associated protein alpha-synuclein. J Biol Chem 286:20710-26