G protein-coupled receptor kinases (GRK) and arrestins act in concert to ensure rapid termination of G protein- mediated signaling by G protein-coupled receptors. Dopaminergic signaling via striatal dopamine receptors play a critical role in controlling multiple forms of behavior in the normal and diseased brain. Abnormalities of signaling by dopamine receptors have been strongly implicated in Parkinson's disease (PD) and motor complications caused by dopamine replacement therapy such as L-DOPA-induced dyskinesia. Dopamine receptors undergo GRK- and arrestin-dependent desensitization, and deficits in this process may underlie signaling abnormalities caused by the loss of dopamine or by dopaminergic drugs in PD. The lentivirus- mediated overexpression of GRK6 in the dopamine-depleted striatum normalizes behavior and dopamine receptor signaling in the rodent and primate animal models of PD, suggesting a critical role for GRKs in the regulation of dopamine receptors in the brain. This project is designed to explore the role of GRKs and arrestins in the dopaminergic regulation of behavior in live animals as well as fine molecular mechanisms of the GRK function in vivo.
First Aim will examine specific roles of GRK isoforms in antagonizing L-DOPA- induced rotations and behavioral sensitization to L-DOPA in hemiparkinsonian rats. We will inject lentiviruses encoding wild type GRK isoforms, mutant GRKs with different functions selectively disabled, or GRK microRNAs to knockdown select isoforms into the dopamine-depleted striatum and test for rotational behavior following repeated L-DOPA administration.
The Aim 2 is designed to test whether simultaneous overexpression of an arrestin and a GRK is more potent than overexpression of the same GRK alone in suppressing L-DOPA-induced motor symptoms.
In Aim 3, we will examine the alterations in down-stream signaling pathways caused by perturbations in the GRK expression or function.
In Aim 4, the feasibility of harnessing the receptor desensitization process to improve therapy in Parkinson's disease will be examined. The results of these experiments will help to sort out specific roles of GRK and arrestin isoforms in regulating dopaminergic signaling in the brain in vivo. The studies will pave the way to development of novel therapeutic approaches to control motor deficits in Parkinsons'disease and complications of L-DOPA therapy.

Public Health Relevance

Parkinson's disease is a neurodegenerative disorder that causes movement problems treated with levodopa, which is an effective drug but looses effectiveness in time and has severe side effects. We do not know what causes problems with levodopa therapy or its produces its side effects. This project is designed to explore whether the proteins called arrestins and G protein-coupled receptor kinases play a role in complications of the levodopa therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS065868-05
Application #
8451473
Study Section
Molecular Neuropharmacology and Signaling Study Section (MNPS)
Program Officer
Sieber, Beth-Anne
Project Start
2009-05-15
Project End
2014-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
5
Fiscal Year
2013
Total Cost
$322,720
Indirect Cost
$115,848
Name
Vanderbilt University Medical Center
Department
Pharmacology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
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Kook, S; Zhan, X; Cleghorn, W M et al. (2014) Caspase-cleaved arrestin-2 and BID cooperatively facilitate cytochrome C release and cell death. Cell Death Differ 21:172-84
Gurevich, Vsevolod V; Gurevich, Eugenia V (2014) Overview of different mechanisms of arrestin-mediated signaling. Curr Protoc Pharmacol 67:Unit 2.10.
Gurevich, Vsevolod V; Gurevich, Eugenia V (2014) Extensive shape shifting underlies functional versatility of arrestins. Curr Opin Cell Biol 27:1-9
Gurevich, Vsevolod V; Gurevich, Eugenia V (2013) Structural determinants of arrestin functions. Prog Mol Biol Transl Sci 118:57-92
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Breitman, Maya; Kook, Seunghyi; Gimenez, Luis E et al. (2012) Silent scaffolds: inhibition OF c-Jun N-terminal kinase 3 activity in cell by dominant-negative arrestin-3 mutant. J Biol Chem 287:19653-64
Gurevich, Eugenia V; Tesmer, John J G; Mushegian, Arcady et al. (2012) G protein-coupled receptor kinases: more than just kinases and not only for GPCRs. Pharmacol Ther 133:40-69
Mushegian, Arcady; Gurevich, Vsevolod V; Gurevich, Eugenia V (2012) The origin and evolution of G protein-coupled receptor kinases. PLoS One 7:e33806

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