A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter

Karl Mattias Rickhag; Freja Herborg Hansen; Gunnar Sørensen; Kristine Nørgaard Strandfelt; Bjørn Andresen; Kamil Gotfryd; Kenneth L Madsen; Ib Vestergaard Klewe; Ina Ammendrup-Johnsen; Jacob Eriksen; Amy H Newman; Ernst-Martin Füchtbauer; Jesus Gomeza; David P D Woldbye; Gitta Wörtwein; Ulrik Gether

doi:10.1038/ncomms2568

A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter

Karl Mattias Rickhag, Freja Herborg Hansen, Gunnar Sørensen, Kristine Nørgaard Strandfelt, Bjørn Andresen, Kamil Gotfryd, Kenneth L Madsen, Ib Vestergaard Klewe, Ina Ammendrup-Johnsen, Jacob Eriksen, Amy H Newman, Ernst-Martin Füchtbauer, Jesus Gomeza, David P D Woldbye, Gitta Wörtwein, Ulrik Gether

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

34 Citations (Scopus)

Abstract

The dopamine transporter mediates reuptake of dopamine from the synaptic cleft. The cellular mechanisms controlling dopamine transporter levels in striatal nerve terminals remain poorly understood. The dopamine transporters contain a C-terminal PDZ (PSD-95/Discs-large/ZO-1) domain-binding sequence believed to bind synaptic scaffolding proteins, but its functional significance is uncertain. Here we demonstrate that two different dopamine transporter knock-in mice with disrupted PDZ-binding motifs (dopamine transporter-AAA and dopamine transporter+Ala) are characterized by dramatic loss of dopamine transporter expression in the striatum, causing hyperlocomotion and attenuated response to amphetamine. In cultured dopaminergic neurons and striatal slices from dopamine transporter-AAA mice, we find markedly reduced dopamine transporter surface levels and evidence for enhanced constitutive internalization. In dopamine transporter-AAA neurons, but not in wild-type neurons, surface levels are rescued in part by expression of a dominant-negative dynamin mutation (K44A). Our findings suggest that PDZ-domain interactions are critical for synaptic distribution of dopamine transporter in vivo and thereby for proper maintenance of dopamine homoeostasis.

Original language	English
Article number	1580
Journal	Nature Communications
Volume	4
Number of pages	14
ISSN	2041-1723
DOIs	https://doi.org/10.1038/ncomms2568
Publication status	Published - 12 Mar 2013

Keywords

Amino Acid Sequence
Amphetamine
Animals
Behavior, Animal
Binding Sites
Biological Transport
Carrier Proteins
Dopamine
Dopamine Plasma Membrane Transport Proteins
Dopaminergic Neurons
Endocytosis
Endoplasmic Reticulum
Immunohistochemistry
Locomotion
Mice
Mice, Knockout
Mutation
Neostriatum
Nuclear Proteins
PDZ Domains
Phenotype
Presynaptic Terminals
Protein Binding
Structure-Activity Relationship

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Rickhag, K. M., Hansen, F. H., Sørensen, G., Strandfelt, K. N., Andresen, B., Gotfryd, K., Madsen, K. L., Klewe, I. V., Ammendrup-Johnsen, I., Eriksen, J., Newman, A. H., Füchtbauer, E.-M., Gomeza, J., Woldbye, D. P. D., Wörtwein, G., & Gether, U. (2013). A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter. Nature Communications, 4, Article 1580. https://doi.org/10.1038/ncomms2568

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title = "A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter",

abstract = "The dopamine transporter mediates reuptake of dopamine from the synaptic cleft. The cellular mechanisms controlling dopamine transporter levels in striatal nerve terminals remain poorly understood. The dopamine transporters contain a C-terminal PDZ (PSD-95/Discs-large/ZO-1) domain-binding sequence believed to bind synaptic scaffolding proteins, but its functional significance is uncertain. Here we demonstrate that two different dopamine transporter knock-in mice with disrupted PDZ-binding motifs (dopamine transporter-AAA and dopamine transporter+Ala) are characterized by dramatic loss of dopamine transporter expression in the striatum, causing hyperlocomotion and attenuated response to amphetamine. In cultured dopaminergic neurons and striatal slices from dopamine transporter-AAA mice, we find markedly reduced dopamine transporter surface levels and evidence for enhanced constitutive internalization. In dopamine transporter-AAA neurons, but not in wild-type neurons, surface levels are rescued in part by expression of a dominant-negative dynamin mutation (K44A). Our findings suggest that PDZ-domain interactions are critical for synaptic distribution of dopamine transporter in vivo and thereby for proper maintenance of dopamine homoeostasis.",

keywords = "Amino Acid Sequence, Amphetamine, Animals, Behavior, Animal, Binding Sites, Biological Transport, Carrier Proteins, Dopamine, Dopamine Plasma Membrane Transport Proteins, Dopaminergic Neurons, Endocytosis, Endoplasmic Reticulum, Immunohistochemistry, Locomotion, Mice, Mice, Knockout, Mutation, Neostriatum, Nuclear Proteins, PDZ Domains, Phenotype, Presynaptic Terminals, Protein Binding, Structure-Activity Relationship",

author = "Rickhag, {Karl Mattias} and Hansen, {Freja Herborg} and Gunnar S{\o}rensen and Strandfelt, {Kristine N{\o}rgaard} and Bj{\o}rn Andresen and Kamil Gotfryd and Madsen, {Kenneth L} and Klewe, {Ib Vestergaard} and Ina Ammendrup-Johnsen and Jacob Eriksen and Newman, {Amy H} and Ernst-Martin F{\"u}chtbauer and Jesus Gomeza and Woldbye, {David P D} and Gitta W{\"o}rtwein and Ulrik Gether",

year = "2013",

month = mar,

day = "12",

doi = "10.1038/ncomms2568",

language = "English",

volume = "4",

journal = "Nature Communications",

issn = "2041-1723",

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Rickhag, KM, Hansen, FH, Sørensen, G, Strandfelt, KN, Andresen, B, Gotfryd, K, Madsen, KL, Klewe, IV, Ammendrup-Johnsen, I, Eriksen, J, Newman, AH, Füchtbauer, E-M, Gomeza, J, Woldbye, DPD, Wörtwein, G & Gether, U 2013, 'A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter', Nature Communications, vol. 4, 1580. https://doi.org/10.1038/ncomms2568

A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter. / Rickhag, Karl Mattias; Hansen, Freja Herborg; Sørensen, Gunnar et al.
In: Nature Communications, Vol. 4, 1580, 12.03.2013.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter

AU - Rickhag, Karl Mattias

AU - Hansen, Freja Herborg

AU - Sørensen, Gunnar

AU - Strandfelt, Kristine Nørgaard

AU - Andresen, Bjørn

AU - Gotfryd, Kamil

AU - Madsen, Kenneth L

AU - Klewe, Ib Vestergaard

AU - Ammendrup-Johnsen, Ina

AU - Eriksen, Jacob

AU - Newman, Amy H

AU - Füchtbauer, Ernst-Martin

AU - Gomeza, Jesus

AU - Woldbye, David P D

AU - Wörtwein, Gitta

AU - Gether, Ulrik

PY - 2013/3/12

Y1 - 2013/3/12

N2 - The dopamine transporter mediates reuptake of dopamine from the synaptic cleft. The cellular mechanisms controlling dopamine transporter levels in striatal nerve terminals remain poorly understood. The dopamine transporters contain a C-terminal PDZ (PSD-95/Discs-large/ZO-1) domain-binding sequence believed to bind synaptic scaffolding proteins, but its functional significance is uncertain. Here we demonstrate that two different dopamine transporter knock-in mice with disrupted PDZ-binding motifs (dopamine transporter-AAA and dopamine transporter+Ala) are characterized by dramatic loss of dopamine transporter expression in the striatum, causing hyperlocomotion and attenuated response to amphetamine. In cultured dopaminergic neurons and striatal slices from dopamine transporter-AAA mice, we find markedly reduced dopamine transporter surface levels and evidence for enhanced constitutive internalization. In dopamine transporter-AAA neurons, but not in wild-type neurons, surface levels are rescued in part by expression of a dominant-negative dynamin mutation (K44A). Our findings suggest that PDZ-domain interactions are critical for synaptic distribution of dopamine transporter in vivo and thereby for proper maintenance of dopamine homoeostasis.

AB - The dopamine transporter mediates reuptake of dopamine from the synaptic cleft. The cellular mechanisms controlling dopamine transporter levels in striatal nerve terminals remain poorly understood. The dopamine transporters contain a C-terminal PDZ (PSD-95/Discs-large/ZO-1) domain-binding sequence believed to bind synaptic scaffolding proteins, but its functional significance is uncertain. Here we demonstrate that two different dopamine transporter knock-in mice with disrupted PDZ-binding motifs (dopamine transporter-AAA and dopamine transporter+Ala) are characterized by dramatic loss of dopamine transporter expression in the striatum, causing hyperlocomotion and attenuated response to amphetamine. In cultured dopaminergic neurons and striatal slices from dopamine transporter-AAA mice, we find markedly reduced dopamine transporter surface levels and evidence for enhanced constitutive internalization. In dopamine transporter-AAA neurons, but not in wild-type neurons, surface levels are rescued in part by expression of a dominant-negative dynamin mutation (K44A). Our findings suggest that PDZ-domain interactions are critical for synaptic distribution of dopamine transporter in vivo and thereby for proper maintenance of dopamine homoeostasis.

KW - Amino Acid Sequence

KW - Amphetamine

KW - Animals

KW - Behavior, Animal

KW - Binding Sites

KW - Biological Transport

KW - Carrier Proteins

KW - Dopamine

KW - Dopamine Plasma Membrane Transport Proteins

KW - Dopaminergic Neurons

KW - Endocytosis

KW - Endoplasmic Reticulum

KW - Immunohistochemistry

KW - Locomotion

KW - Mice

KW - Mice, Knockout

KW - Mutation

KW - Neostriatum

KW - Nuclear Proteins

KW - PDZ Domains

KW - Phenotype

KW - Presynaptic Terminals

KW - Protein Binding

KW - Structure-Activity Relationship

U2 - 10.1038/ncomms2568

DO - 10.1038/ncomms2568

M3 - Journal article

C2 - 23481388

SN - 2041-1723

VL - 4

JO - Nature Communications

JF - Nature Communications

M1 - 1580

ER -

A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter

Abstract

Keywords

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