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Rin GTPase And Dopamine Transporter Trafficking: Linking Mechanisms To Behavior

Rin GTP 酶和多巴胺转运蛋白贩运：将机制与行为联系起来

基本信息

批准号：
9267953
负责人：
Carolyn Gray Sweeney
金额：
$ 3.06万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-10 至 2018-04-09
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9267953
关键词：
Acute Address Amphetamines Antidepressive Agents Attention deficit hyperactivity disorder Behavior Behavioral Binding Binding Sites Brain Bupropion Carrier Proteins Cell Line Cell Surface Proteins Cell membrane Cell surface Cells Chimera organism Chimeric Proteins Chronic Co-Immunoprecipitations Cocaine Cognition Complex Corpus striatum structure County Data Disease Dopamine Dopaminergic Cell Down-Regulation Drug Prescriptions Endocytosis Energy Transfer Extracellular Space Foundations Guanosine Triphosphate Phosphohydrolases In Situ Investigation Laboratories Lead Link LoxP-flanked allele Mediating Methylphenidate Microscopy Molecular Molecular Target Motivation Motor Activity Movement Mus N-terminal Neurobiology Neurons Neurosciences Parkinson Disease Pharmaceutical Preparations Physiological Preparation Process Property Protein Kinase C Psychotropic Drugs Regulation Reporting Rewards Ritalin Role Schizophrenia Scientist Signal Transduction Slice Surface Synapses Testing Therapeutic Therapeutic Agents Time Training Ubiquitin Ubiquitination United States addiction career dopamine transporter drug of abuse extracellular functional loss in vivo knock-down mutant novel therapeutic intervention presynaptic psychostimulant public health relevance response small hairpin RNA stimulant abuse synergism trafficking transmission process

项目摘要

DESCRIPTION (provided by applicant): Dopamine (DA) signaling in the CNS is essential for modulating complex behaviors such as movement, cognition, reward, and motivation. Aberrant DAergic transmission is directly linked to Parkinson's disease, attention-deficit hyperactivity disorder, schizophrenia and addiction. The plasma membrane dopamine transporter (DAT) is central to DAergic signaling, and both limits extracellular DA availability and maintains presynaptic DA stores. Importantly, DAT is the primary target for both addictive and therapeutic psychoactive drugs, such as cocaine, amphetamine, methylphenidate (Ritalin), and the antidepressant bupropion (Wellbutrin), all of which competitively inhibit DAT activity. A wealth of data supports the premise that DAT constitutively traffics to and from the cell surface, and that protein kinase C (PKC) activation rapidly decreases DAT cell surface expression by acutely modulating DAT trafficking rates. While some progress has been made investigating the molecular mechanisms that govern regulated DAT trafficking, a detailed understanding of this complex process has yet to be achieved. Further, the physiological relevance that DAT trafficking imposes onto DAergic function is poorly understood. Our laboratory reported that neuronal GTPase, Rin (RIT2), specifically binds to the DAT carboxy terminus and is required for PKC-stimulated DAT internalization. In the proposed studies, I plan to investigate the molecular underpinnings required for DAT/Rin interactions, and how these interactions potentially mediate synergy between cytosolic DAT domains. Aim 1 studies will use co-immunoprecipitations and FRET microscopy approaches with chimeric DAT proteins to define the intracellular DAT domains that are necessary and sufficient to confer DAT/Rin interactions. Chimeric and point mutant transporters will further reveal the potential synergistic requirement of DAT amino- and carboxy-termini for PKC-stimulated DAT internalization. Aim 2 studies will directly test the potential role of Rin-mediated DAT trafficking in DAergic behaviors in vivo, using cell-specific, AAV2-mediated Rin knockdown in DAergic terminal regions. Taken together, these studies will provide the first investigations that directly examine how DAT trafficking potentially impacts rewarding behavior. Moreover, completion of this investigative line will provide me with outstanding training in molecular and behavioral neuroscience.