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Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function

Extracellular dopamine (DA) levels are constrained by the presynaptic DA transporter (DAT), a major psychostimulant target. Despite its necessity for DA neurotransmission, DAT regulation in situ is poorly understood, and it is unknown whether regulated DAT trafficking impacts dopaminergic signaling...

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Autores principales: Kearney, Patrick J., Bolden, Nicholas C., Kahuno, Elizabeth, Conklin, Tucker L., Martin, Gilles E., Lubec, Gert, Melikian, Haley E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9943899/
https://www.ncbi.nlm.nih.gov/pubmed/36640864
http://dx.doi.org/10.1016/j.jbc.2023.102900
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author Kearney, Patrick J.
Bolden, Nicholas C.
Kahuno, Elizabeth
Conklin, Tucker L.
Martin, Gilles E.
Lubec, Gert
Melikian, Haley E.
author_facet Kearney, Patrick J.
Bolden, Nicholas C.
Kahuno, Elizabeth
Conklin, Tucker L.
Martin, Gilles E.
Lubec, Gert
Melikian, Haley E.
author_sort Kearney, Patrick J.
collection PubMed
description Extracellular dopamine (DA) levels are constrained by the presynaptic DA transporter (DAT), a major psychostimulant target. Despite its necessity for DA neurotransmission, DAT regulation in situ is poorly understood, and it is unknown whether regulated DAT trafficking impacts dopaminergic signaling and/or behaviors. Leveraging chemogenetics and conditional gene silencing, we found that activating presynaptic Gq-coupled receptors, either hM3Dq or mGlu5, drove rapid biphasic DAT membrane trafficking in ex vivo striatal slices, with region-specific differences between ventral and dorsal striata. DAT insertion required D2 DA autoreceptors and intact retromer, whereas DAT retrieval required PKC activation and Rit2. Ex vivo voltammetric studies revealed that DAT trafficking impacts DA clearance. Furthermore, dopaminergic mGlu5 silencing elevated DAT surface expression and abolished motor learning, which was rescued by inhibiting DAT with a subthreshold CE-158 dose. We discovered that presynaptic DAT trafficking is complex, multimodal, and region specific, and for the first time, we identified cell autonomous mechanisms that govern presynaptic DAT tone. Importantly, the findings are consistent with a role for regulated DAT trafficking in DA clearance and motor function.
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spelling pubmed-99438992023-02-23 Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function Kearney, Patrick J. Bolden, Nicholas C. Kahuno, Elizabeth Conklin, Tucker L. Martin, Gilles E. Lubec, Gert Melikian, Haley E. J Biol Chem Research Article Extracellular dopamine (DA) levels are constrained by the presynaptic DA transporter (DAT), a major psychostimulant target. Despite its necessity for DA neurotransmission, DAT regulation in situ is poorly understood, and it is unknown whether regulated DAT trafficking impacts dopaminergic signaling and/or behaviors. Leveraging chemogenetics and conditional gene silencing, we found that activating presynaptic Gq-coupled receptors, either hM3Dq or mGlu5, drove rapid biphasic DAT membrane trafficking in ex vivo striatal slices, with region-specific differences between ventral and dorsal striata. DAT insertion required D2 DA autoreceptors and intact retromer, whereas DAT retrieval required PKC activation and Rit2. Ex vivo voltammetric studies revealed that DAT trafficking impacts DA clearance. Furthermore, dopaminergic mGlu5 silencing elevated DAT surface expression and abolished motor learning, which was rescued by inhibiting DAT with a subthreshold CE-158 dose. We discovered that presynaptic DAT trafficking is complex, multimodal, and region specific, and for the first time, we identified cell autonomous mechanisms that govern presynaptic DAT tone. Importantly, the findings are consistent with a role for regulated DAT trafficking in DA clearance and motor function. American Society for Biochemistry and Molecular Biology 2023-01-12 /pmc/articles/PMC9943899/ /pubmed/36640864 http://dx.doi.org/10.1016/j.jbc.2023.102900 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Kearney, Patrick J.
Bolden, Nicholas C.
Kahuno, Elizabeth
Conklin, Tucker L.
Martin, Gilles E.
Lubec, Gert
Melikian, Haley E.
Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function
title Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function
title_full Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function
title_fullStr Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function
title_full_unstemmed Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function
title_short Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function
title_sort presynaptic gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9943899/
https://www.ncbi.nlm.nih.gov/pubmed/36640864
http://dx.doi.org/10.1016/j.jbc.2023.102900
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